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Sample records for biocontrol strain clonostachys

  1. Development of a biocontrol agent for plant disease control with special emphasis on the near commercial fungal antagonist Clonostachys rosea strain "IK726"

    DEFF Research Database (Denmark)

    Jensen, Dan Funck; Knudsen, Inge M.B.; Lübeck, Mette;

    2007-01-01

    . Among the success stories for control of seed- and soilborne diseases are fungal biocontrol agents based on Trichoderma harzianum, Clonostachys rosea and Conithyrium minitans, and bacterial biocontrol agents based on strains of Agrobacterium, Pseudomonas and Streptomyces. We have developed C. rosea...

  2. GUS and GFP transformation of the biocontrol strain Clonostachys rosea IK726 and the use of these marker genes in ecological studies

    DEFF Research Database (Denmark)

    Lübeck, M.; Knudsen, I.M.B.; Jensen, B.;

    2002-01-01

    Marker genes were introduced in the biocontrol strain Clonostachys rosea IK726 (IBT 9371) as a tool for monitoring the strain in ecological studies. The beta-glucuronidase (GUS) reporter gene and a gene encoding the green fluorescent protein (GFP) were, in separate experiments, integrated...... into the genome of IK726 using the Hygromycin B (HygB) resistance gene as selective marker. In order to select GUS and GFP transformants that resembled the wildtype strain, growth rate, production of enzymes and of metabolites of four GUS positive and six GFP positive transformants were tested in vitro....... In addition, the biocontrol efficacy against disease caused by seed-borne Fusarium culmorum was evaluated on barley grown in sand. Compared to the wildtype, two selected GUS and GFP transformants, IK726c5 and IK726d11, did not vary in physiological properties. Both maintained the ability to colonize barley...

  3. Real-time RT-PCR expression analysis of chitinase and endoglucanase genes in the three-way interaction between the biocontrol strain Clonostachys rosea IK726, Botrytis cinera and strawberry

    DEFF Research Database (Denmark)

    Mamarabadi, Mojtaba; Jensen, Birgit; Jensen, Søren Dan Funck;

    2008-01-01

    Clonostachys rosea is a well-known biocontrol agent against Botrytis cinerea, the causal agent of gray mold in strawberry. The activity of cell wall-degrading enzymes might play a significant role for successful biocontrol by C. rosea. The expression pattern of four chitinases, and two endoglucan......Clonostachys rosea is a well-known biocontrol agent against Botrytis cinerea, the causal agent of gray mold in strawberry. The activity of cell wall-degrading enzymes might play a significant role for successful biocontrol by C. rosea. The expression pattern of four chitinases, and two...... endoglucanase genes from C. rosea strain IK726 was analyzed using real-time RT-PCR in vitro and in strawberry leaves during interaction with B. cinerea. Specific primers were designed for ß-tubulin genes from C. rosea and B. cinerea, respectively, and a gene encoding a DNA-binding protein (DBP) from strawberry......, allowing in situ activity assessment of each fungus in vitro and during their interaction on strawberry leaves. Growth of B. cinerea was inhibited in all pathogen-antagonist interactions while the activity of IK726 was slightly increased. In all in vitro interactions, four of the six genes were upregulated...

  4. Transaldolase gene Tal67 enhances the biocontrol activity of Clonostachys rosea 67-1 against Sclerotinia sclerotiorum.

    Science.gov (United States)

    Liu, Jing-Yu; Li, Shi-Dong; Sun, Man-Hong

    2016-06-01

    Clonostachys rosea is a promising biocontrol agent that parasitizes various fungal plant pathogens. In this paper, transaldolase gene Tal67 was found to be greatly upregulated in C. rosea isolate 67-1 during mycoparasitism of Sclerotinia sclerotiorum sclerotia. Quantitative real-time PCR revealed a significant increase in expression at 0-48 h after induction by sclerotia, and the level peaked at 13.9-fold higher than the control at 24 h. Gene disruption led to a decrease in the growth rate of the Tal67-deficient strain ΔTal67 to 5.3 mm/day, which was much lower than the wild type and the complemented strain ΔTal67+ (P Sclerotinia stem rot was evaluated in the greenhouse, and the control efficiency of isolate 67-1 reached 65.3%, while the efficiency of the ΔTal67 strain decreased sharply to 17.8%, and the complemented strain ΔTal67+ recovered to 64.8%. These results suggest that Tal67 plays an important role in the growth and biocontrol activity of C. rosea. PMID:27130824

  5. Evaluation of Trichoderma spp. and Clonostachys spp. Strains to Control Fusarium circinatum in Pinus radiata Seedlings Evaluación de Cepas de Trichoderma spp. y Clonostachys spp. para Controlar Fusarium circinatum en Plántulas de Pinus radiata

    Directory of Open Access Journals (Sweden)

    Priscila Moraga-Suazo

    2011-09-01

    Full Text Available The fungus Fusarium circinatum Nirenberg & O’Donnell causes pine pitch canker, an important disease for conifers worldwide. F. circinatum was first detected in Chile in 2001 and to date is present in nurseries and clonal hedges from Libertador General Bernardo O’Higgins Region to Los Rios Region. The purpose of this study was to evaluate the potential of Trichoderma spp. and Clonostachys spp. strains to control F. circinatum in Pinus radiata D. Don seedlings in the absence of other effective control methods. Eighty-one Trichoderma spp. and Clonostachys spp. strains were evaluated through in vitro assays to determine their ability to act as antagonists of F. circinatum and 21 strains were tested for their ability to reduce post-emergence mortality and increase P. radiata survival under greenhouse conditions. During in vitro experiments, 15 strains of Trichoderma inhibited mycelial growth of the pathogen by more than 60% and one strain of Clonostachys showed parasitism of F. circinatum hyphae. Greenhouse experiments showed no control of the disease when the antagonists were added to substrate after the pathogen. However, when the antagonists were added before the pathogen, four strains (Clonostachys UDC-32 and UDC-222 and Trichoderma UDC-23 and UDC-408 reduced post-emergence mortality between 80 and 100%. Among these strains, only Clonostachys UDC-222 significantly increased the survival of P. radiata seedlings. These results showed that Clonostachys UDC-222 has the potential to be used as a biocontrol agent against F. circinatum in the production of P. radiata plants.Fusarium circinatum Nirenberg & O’Donnell es el hongo que causa el cancro resinoso del pino, una enfermedad de importancia mundial en coníferas. En Chile, F. cicirnatum fue detectado por primera vez el año 2001 y a la fecha se encuentra presente en algunos viveros y huertos clonales desde la Región del Libertador General Bernardo O’Higgins hasta la Región de Los R

  6. Towards understanding the ecology and mechanisms of biocontrol of Clonostachys rosea IK726

    Institute of Scientific and Technical Information of China (English)

    Mette Lübeck; Inge M B Knudsen; Birgit Jensen; Mojtaba Mamarabadi; Dan Funck Jensen

    2004-01-01

    @@ Clonostachys rosea (syn. Gliocladium roseum ) IK726 was originally selected as an effective biocontrol agent (BCA) against cereal seed borne diseases caused by Fusarium culmorum and Bipolaris sorokiniana. We have studied the efficacy of the antagonist against different pathogens in several crops and found that the antagonist also is able to control Alternaria radicina and A. dauci on carrot seeds and different cold-storage fungi in acorns. IK726 is also able to reduce severity of soil borne Pythium spp. in cabbage, carrot and sugar beet. In addition, growth-promoting effects of IK726 have been demonstrated in barley and tomato. In order to develop and improve application methods and control strategies, essential basic studies of ecology and the mechanisms of control of IK726 is needed and has led us to use various molecular tools. The UP-PCR technology is used for strain recognition and we have developed GUS and GFP-transformants that resembles the wildtype strain in ecological fitness parameters. Using either the GUS-transformant or UP-PCR we have found that IK726, when applied with seeds, reproduces and survives several months in the rhizosphere of field grown barley and carrot.The GFP-transformant is used to study the behavior and in situ interactions of the antagonist with pathogens and plants. Using the GFP marker, we have observed conidial germination, colonization and conidiogenesis in natural soil, in vermiculite and on carrot and barley seed and roots and on barley leaves. Moreover in situ interactions with Alternaria on carrot material have been studied. The modes of action of C. rosea are not well understood but enzymatic activity, mycoparasitism, substrate competition, antibiosis and induced resistance are thought to play a role. Barley treated with C. rosea IK726 has an enhanced chitinolytic and glucanolytic activity compared to the activity in non-treated barley in pot experiments with field soil. Identification of chitinases from IK726 and studies

  7. Control of Botrytis cinerea in Eucalyptus globulus Mini-Cuttings Using Clonostachys and Trichoderma Strains Control de Botrytis cinerea en miniestacas de Eucalyptus globulus Utilizando Cepas de Clonostachys y Trichoderma

    Directory of Open Access Journals (Sweden)

    Salomé Zaldúa

    2010-12-01

    Full Text Available Botrytis cinerea Pers. ex Fr. causes the disease known as gray mold in more than 200 hosts. It is one of the most important pathogens in Chilean forest nurseries and Eucalyptus globulus Labill. is one of the most susceptible species, especially in vegetative reproduction systems. Clonostachys and Trichoderma strains were selected as potential biocontrol agents of gray mold in previous research by the authors. The objective of this study was to evaluate the effectiveness of antagonistic fungi to control B. cinerea in E. globulus mini-cuttings. Five fungi strains were tested and applied weekly, two Clonostachys and three Trichoderma (5 x 10(6 conidia mL-1. In addition, comparison treatments were also used: absolute control (water and fungicide application. The experiment was carried out under operational conditions to produce E. globulus mini-cuttings. The Clonostachys UDC-A10 and UDC-A11 strains reduce mini-cutting mortality caused by B. cinerea in 54 and 71%, respectively, and with effects similar to those achieved by fungicides. Clonostachys UDC-A11 reduces the disease progression rate with the same statistical results as fungicides. A negative effect of applying fungicides on rooting of the surviving mini-cuttings was also confirmed. These results demonstrate the effectiveness of Clonostachys as a control agent against gray mold disease in E. globulus mini-cuttings.Botrytis cinerea Pers. ex Fr. ocasiona la enfermedad conocida como moho gris en más de 200 hospederos. En Chile es uno de los patógenos más importantes en viveros forestales, siendo Eucalyptus globulus Labill. una de las especies más susceptibles, especialmente en los sistemas de reproducción vegetativa. En investigaciones previas, realizadas por los autores, se seleccionaron cepas de Clonostachys y Trichoderma como potenciales agentes de biocontrol del moho gris. El objetivo fue evaluar la eficacia de hongos antagonistas en el control de B. cinerea en mini-estacas de E

  8. The ABC transporter ABCG29 is involved in H2O2 tolerance and biocontrol traits in the fungus Clonostachys rosea.

    Science.gov (United States)

    Dubey, Mukesh; Jensen, Dan Funck; Karlsson, Magnus

    2016-04-01

    For successful biocontrol interactions, biological control organisms must tolerate toxic metabolites produced by themselves or plant pathogens during mycoparasitic/antagonistic interactions, by host plant during colonization of the plant, and xenobiotics present in the environment. ATP-binding cassette (ABC) transporters can play a significant role in tolerance of toxic compounds by mediating active transport across the cellular membrane. This paper reports on functional characterization of an ABC transporter ABCG29 in the biocontrol fungus Clonostachys rosea strain IK726. Gene expression analysis showed induced expression of abcG29 during exposure to the Fusarium spp. mycotoxin zearalenone (ZEA) and the fungicides Cantus, Chipco Green and Apron. Expression of abcG29 in C. rosea was significantly higher during C. rosea-C. rosea (Cr-Cr) interaction or in exposure to C. rosea culture filtrate for 2 h, compared to interaction with Fusarium graminearum or 2 h exposure to F. graminearum culture filtrate. In contrast with gene expression data, ΔabcG29 strains did not display reduced tolerance towards ZEA, fungicides or chemical agents known for inducing oxidative, cell wall or osmotic stress, compared to C. rosea WT. The exception was a significant reduction in tolerance to H2O2 (10 mM) in ΔabcG29 strains when conidia were used as an inoculum. The antagonistic ability of ΔabcG29 strains towards F. graminearum, Fusarium oxysporum or Botrytis cinerea in dual plate assays were not different compared with WT. However, in biocontrol assays ΔabcG29 strains displayed reduced ability to protect Arabidopsis thaliana leaves from B. cinerea, and barley seedling from F. graminearum as measured by an A. thaliana detached leaf assay and a barley foot rot disease assay, respectively. These data show that the ABCG29 is dispensable for ZEA and fungicides tolerance, and antagonism but not H2O2 tolerance and biocontrol effects in C. rosea. PMID:26520102

  9. Draft Genome Sequence of Mycoparasite Clonostachys rosea Strain 67-1

    OpenAIRE

    Sun, Zhan-Bin; Sun, Man-Hong; Li, Shi-Dong

    2015-01-01

    Clonostachys rosea is a promising mycoparasite. In this study, we sequenced the draft genome of the highly effective strain 67-1 using the Illumina HiSeq 2500 sequencing platform. The genome is 55.4 Mb with a G+C content of 49.2% and provides a powerful resource for future studies on the molecular mechanisms underlying Clonostachys rosea’s antagonism on fungal pathogens.

  10. Three encochitinase-encoding genes identified in the biocontrol fungus Clonostachys rosea are differentially expressed

    DEFF Research Database (Denmark)

    Mamarabadi, Mojtaba; Jensen, Birgit; Lübeck, Mette

    2008-01-01

    . rosea were made by disruption of the endochitinase genes via Agrobacterium-mediated transformation and their biocontrol activity was evaluated. While in planta bioassays showed no significant difference in biocontrol efficacy between the disruptants and the wildtype, the real time RT-PCR analysis showed...

  11. [Improvement of Trichoderma strains for biocontrol].

    Science.gov (United States)

    Benítez, T; Rey, M; Delgado-Jarana, J; Rincón, A M; Limón, M C

    2000-03-01

    The use of the fungal genus Trichoderma to control fungal plant diseases is a promising alternative to the use of chemical compounds. The aim of this work has been to obtain Trichoderma strains with improved capacity as biological control agents. To do so, the hydrolytic capacity on fungal cell walls of strains of the fungus Trichoderma harzianum has been increased. On one hand, transformation experiments with genes which coded for chitinases and glucanases have been carried out in T. harzianumstra ins. On the other hand, the medium composition has also been modified in order to eliminate proteolytic degradation of some of the overproduced enzymes. Finally, hybrid chitinolytic enzymes with substrate-binding domains have been produced as an alternative to obtain improved biocontrol strains. The transformant strains, when compared with the wild type, showed improved antifungal capacity against the phytopathogenic fungus Rhizoctonia solani, in in vitro experiments. PMID:15762779

  12. Biological Control of Pathogens Causing Root Rot Complex in Field Pea Using Clonostachys rosea Strain ACM941.

    Science.gov (United States)

    Xue, Allen G

    2003-03-01

    ABSTRACT Pea root rot complex (PRRC), caused by Alternaria alternata, Aphanomyces euteiches, Fusarium oxysporum f. sp. pisi, F. solani f. sp. pisi, Mycosphaerella pinodes, Pythium spp., Rhizoctonia solani, and Sclerotinia sclerotiorum, is a major yield-limiting factor for field pea production in Canada. A strain of Clonostachys rosea (syn. Gliocladium roseum), ACM941 (ATCC 74447), was identified as a mycoparasite against these pathogens. When grown near the pathogen, ACM941 often was stimulated to produce lateral branches that grew directly toward the pathogen mycelium, typically entwining around the pathogen mycelium. When applied to the seed, ACM941 propagated in the rhizosphere and colonized the seed coat, hypocotyl, and roots as the plant developed and grew. ACM941 significantly reduced the recovery of all fungal pathogens from infected seed, increased in vitro seed germination by 44% and seedling emergence by 22%, and reduced root rot severity by 76%. The effects were similar to those of thiram fungicide, which increased germination and emergence by 33 and 29%, respectively, and reduced root rot severity by 65%. When soil was inoculated with selected PRRC pathogens in a controlled environment, seed treatment with ACM941 significantly increased emergence by 26, 38, 28, 13, and 21% for F. oxysporum f. sp. pisi, F. solani f. sp. pisi, M. pinodes, R. solani, and S. sclerotiorum, respectively. Under field conditions from 1995 to 1997, ACM941 increased emergence by 17, 23, 22, 13, and 18% and yield by 15, 6, 28, 6, and 19% for the five respective pathogens. The seed treatment effects of ACM941 on these PRRC pathogens were greater or statistically equivalent to those achieved with thiram. Results of this study suggest that ACM941 is an effective bioagent in controlling PRRC and is an alternative to existing chemical products. PMID:18944343

  13. Genome Sequence of the Biocontrol Strain Pseudomonas fluorescens F113

    OpenAIRE

    Redondo-Nieto, M.; M. Barret; Morrisey, J; Germaine, K.; Martínez-Granero, F.; Barahona, E.; Navazo, A.; Sánchez-Contreras, M.; Moynihan, J.; Giddens, S.; Coppoolse, E.; Muriel, C.; Stiekema, W.; Rainey, P; Dowling, D

    2012-01-01

    Pseudomonas fluorescens F113 is a plant growth-promoting rhizobacterium (PGPR) that has biocontrol activity against fungal plant pathogens and is a model for rhizosphere colonization. Here, we present its complete genome sequence, which shows that besides a core genome very similar to those of other strains sequenced within this species, F113 possesses a wide array of genes encoding specialized functions for thriving in the rhizosphere and interacting with eukaryotic organisms.

  14. Complete Genome Sequence of Biocontrol Strain Pseudomonas fluorescens LBUM 223

    OpenAIRE

    Roquigny, Roxane; Arseneault, Tanya; Gadkar, Vijay J.; Novinscak, Amy; Joly, David L.; Filion, Martin

    2015-01-01

    Pseudomonas fluorescens LBUM 223 is a plant growth-promoting rhizobacterium (PGPR) with biocontrol activity against various plant pathogens. It produces the antimicrobial metabolite phenazine-1-carboxylic acid, which is involved in the biocontrol of Streptomyces scabies, the causal agent of common scab of potato. Here, we report the complete genome sequence of P. fluorescens LBUM 223.

  15. Draft genome sequences of the Pseudomonas fluorescens biocontrol strains Wayne1R and Wood1R.

    Science.gov (United States)

    Rong, Xiaoqing; Gurel, Fulya Baysal; Meulia, Tea; McSpadden Gardener, Brian B

    2012-02-01

    Pseudomonas fluorescens strains Wayne1R and Wood1R have proven capacities to improve plant health. Here we report the draft genome sequences and automatic annotations of both strains. Genome comparisons reveal similarities with P. fluorescens strain Pf-5, reveal the novelty of Wood1R, and indicate some genes that may be related to biocontrol.

  16. Draft Genome Sequences of the Pseudomonas fluorescens Biocontrol Strains Wayne1R and Wood1R

    OpenAIRE

    Rong, Xiaoqing; Baysal Gurel, Fulya; Meulia, Tea; McSpadden Gardener, Brian B

    2012-01-01

    Pseudomonas fluorescens strains Wayne1R and Wood1R have proven capacities to improve plant health. Here we report the draft genome sequences and automatic annotations of both strains. Genome comparisons reveal similarities with P. fluorescens strain Pf-5, reveal the novelty of Wood1R, and indicate some genes that may be related to biocontrol.

  17. Assessment of biocontrol strains for reduction of mycotoxins (aflatoxin and CPA) in maize

    Science.gov (United States)

    Aspergillus flavus strains K49, NRRL 21882 (from Afla-Guard®) and AF36 are being developed as biocontrol agents for the control of aflatoxin in maize. In this study, the three non-aflatoxigenic strains were compared to evaluate which is most effective in reducing aflatoxin. Also, we tested these st...

  18. Draft Genome Sequence of a Biocontrol Rhizobacterium, Chryseobacterium kwangjuense Strain KJ1R5, Isolated from Pepper (Capsicum annuum).

    Science.gov (United States)

    Jeong, Jin-Ju; Park, Hongjae; Park, Byeong Hyeok; Mannaa, Mohamed; Sang, Mee Kyung; Choi, In-Geol; Kim, Ki Deok

    2016-01-01

    Strain KJ1R5 of the rhizobacterium ITALIC! Chryseobacterium kwangjuenseis an effective biocontrol agent against Phytophthora blight of pepper caused by a destructive soilborne oomycete, ITALIC! Phytophthora capsici Here, we present the draft genome sequence of strain KJ1R5, which contains genes related to biocontrol, plant growth promotion, and environmental stress adaptation. PMID:27103726

  19. Study on Attenuation Characteristics of Biocontrol Strain Anti-8098A, Bacillus cereus, against Ralstoniasolanacearum

    Institute of Scientific and Technical Information of China (English)

    BoLIU; Ying-ZhiLIN; Yu-JingZHU; Ci-BinGE; YiCAO

    2004-01-01

    The present study dealt with the attenuation characteristics of bacterial-wilt-disease biocontrol strain Anti-8098A, Bacillus cereus, againstpathogeny Ralstonia solanacearum (RS). In order to distinguish the pathogenicity of RS, the attenuation index (radius of the center red ring/radius of the whole mycelium ring, on TTC culture medium) was established (Hayward, 1976), companying with the mortality of tomato

  20. Evaluation of recycled bioplastic pellets and a sprayable formulation for application of an Aspergillus flavus biocontrol strain

    Science.gov (United States)

    Biocontrol of Aspergillus flavus using inoculated bioplastic granules has been proven to be effective under laboratory and field conditions. In the present study, the use of low-density pellets from recycled bioplastic as a biocontrol strain carrier was evaluated. Applying recycled bioplastic pell...

  1. Isolation of Dickeya dadantii strains from potato disease and biocontrol by their bacteriophages.

    Science.gov (United States)

    Soleimani-Delfan, Abbas; Etemadifar, Zahra; Emtiazi, Giti; Bouzari, Majid

    2015-01-01

    One of the most economically important bacterial pathogens of plants and plant products is Dickeya dadantii. This bacterium causes soft rot disease in tubers and other parts of the potato and other plants of the Solanaceae family. The application of restricted host range bacteriophages as biocontrol agents has recently gained widespread interest. This study purposed to isolate the infectious agent of the potato and evaluate its biocontrol by bacteriophages. Two phytopathogenic strains were isolated from infected potatoes, identified based on biochemical and 16S rRNA gene sequencing, and submitted to GenBank as D. dadantii strain pis3 (accession no. HQ423668) and D. dadantii strain sip4 (accession no. HQ423669). Their bacteriophages were isolated from Caspian Sea water by enriching the water filtrate with D. dadantii strains as hosts using spot or overlay methods. On the basis of morphotypes, the isolated bacteriophages were identified as members of the Myoviridae and Siphoviridae families and could inhibit the growth of antibiotic resistant D. dadantii strains in culture medium. Moreover, in Dickeya infected plants treated with bacteriophage, no disease progression was detected. No significant difference was seen between phage-treated and control plants. Thus, isolated bacteriophages can be suggested for the biocontrol of plant disease caused by Dickeya strains.

  2. Complete Genome Sequence of the Endophytic Biocontrol Strain Bacillus velezensis CC09

    Science.gov (United States)

    Cai, Xunchao; Kang, Xingxing; Xi, Huan

    2016-01-01

    Bacillus velezensis is a heterotypic synonym of B. methylotrophicus, B. amyloliquefaciens subsp. plantarum, and Bacillus oryzicola, and has been used to control plant fungal diseases. In order to fully understand the genetic basis of antimicrobial capacities, we did a complete genome sequencing of the endophytic B. velezensis strain CC09. Genes tightly associated with biocontrol ability, including nonribosomal peptide synthetases, polyketide synthetases, iron acquisition, colonization, and volatile organic compound synthesis were identified in the genome. PMID:27688342

  3. Complete Genome Sequence of the Endophytic Biocontrol Strain Bacillus velezensis CC09.

    Science.gov (United States)

    Cai, Xunchao; Kang, Xingxing; Xi, Huan; Liu, Changhong; Xue, Yarong

    2016-01-01

    Bacillus velezensis is a heterotypic synonym of B. methylotrophicus, B. amyloliquefaciens subsp. plantarum, and Bacillus oryzicola, and has been used to control plant fungal diseases. In order to fully understand the genetic basis of antimicrobial capacities, we did a complete genome sequencing of the endophytic B. velezensis strain CC09. Genes tightly associated with biocontrol ability, including nonribosomal peptide synthetases, polyketide synthetases, iron acquisition, colonization, and volatile organic compound synthesis were identified in the genome. PMID:27688342

  4. Characterization of the biocontrol activity of pseudomonas fluorescens strain X reveals novel genes regulated by glucose.

    Directory of Open Access Journals (Sweden)

    Gerasimos F Kremmydas

    Full Text Available Pseudomonas fluorescens strain X, a bacterial isolate from the rhizosphere of bean seedlings, has the ability to suppress damping-off caused by the oomycete Pythium ultimum. To determine the genes controlling the biocontrol activity of strain X, transposon mutagenesis, sequencing and complementation was performed. Results indicate that, biocontrol ability of this isolate is attributed to gcd gene encoding glucose dehydrogenase, genes encoding its co-enzyme pyrroloquinoline quinone (PQQ, and two genes (sup5 and sup6 which seem to be organized in a putative operon. This operon (named supX consists of five genes, one of which encodes a non-ribosomal peptide synthase. A unique binding site for a GntR-type transcriptional factor is localized upstream of the supX putative operon. Synteny comparison of the genes in supX revealed that they are common in the genus Pseudomonas, but with a low degree of similarity. supX shows high similarity only to the mangotoxin operon of Ps. syringae pv. syringae UMAF0158. Quantitative real-time PCR analysis indicated that transcription of supX is strongly reduced in the gcd and PQQ-minus mutants of Ps. fluorescens strain X. On the contrary, transcription of supX in the wild type is enhanced by glucose and transcription levels that appear to be higher during the stationary phase. Gcd, which uses PQQ as a cofactor, catalyses the oxidation of glucose to gluconic acid, which controls the activity of the GntR family of transcriptional factors. The genes in the supX putative operon have not been implicated before in the biocontrol of plant pathogens by pseudomonads. They are involved in the biosynthesis of an antimicrobial compound by Ps. fluorescens strain X and their transcription is controlled by glucose, possibly through the activity of a GntR-type transcriptional factor binding upstream of this putative operon.

  5. Characterization of the biocontrol activity of pseudomonas fluorescens strain X reveals novel genes regulated by glucose.

    Science.gov (United States)

    Kremmydas, Gerasimos F; Tampakaki, Anastasia P; Georgakopoulos, Dimitrios G

    2013-01-01

    Pseudomonas fluorescens strain X, a bacterial isolate from the rhizosphere of bean seedlings, has the ability to suppress damping-off caused by the oomycete Pythium ultimum. To determine the genes controlling the biocontrol activity of strain X, transposon mutagenesis, sequencing and complementation was performed. Results indicate that, biocontrol ability of this isolate is attributed to gcd gene encoding glucose dehydrogenase, genes encoding its co-enzyme pyrroloquinoline quinone (PQQ), and two genes (sup5 and sup6) which seem to be organized in a putative operon. This operon (named supX) consists of five genes, one of which encodes a non-ribosomal peptide synthase. A unique binding site for a GntR-type transcriptional factor is localized upstream of the supX putative operon. Synteny comparison of the genes in supX revealed that they are common in the genus Pseudomonas, but with a low degree of similarity. supX shows high similarity only to the mangotoxin operon of Ps. syringae pv. syringae UMAF0158. Quantitative real-time PCR analysis indicated that transcription of supX is strongly reduced in the gcd and PQQ-minus mutants of Ps. fluorescens strain X. On the contrary, transcription of supX in the wild type is enhanced by glucose and transcription levels that appear to be higher during the stationary phase. Gcd, which uses PQQ as a cofactor, catalyses the oxidation of glucose to gluconic acid, which controls the activity of the GntR family of transcriptional factors. The genes in the supX putative operon have not been implicated before in the biocontrol of plant pathogens by pseudomonads. They are involved in the biosynthesis of an antimicrobial compound by Ps. fluorescens strain X and their transcription is controlled by glucose, possibly through the activity of a GntR-type transcriptional factor binding upstream of this putative operon.

  6. Effects of Biocontrol Strain E26 on Some Ecological Factors

    Institute of Scientific and Technical Information of China (English)

    WANG Hui-min; SUN Yan-li; WANG Jian-hui

    2002-01-01

    This study was to evaluate the ecological risk of strain E26 (Agrobacterium sp. ) by detecting its survival in waters and its effects on rhizosphere microorganisms. The data showed that E26 could not be detected in distilled water, tap water, river water, and rainwater after 36, 36, 8, and 9 days, respectively. E26 did not reveal significant effects on the population of bacteria, fungi, and actinomyces in rhizosphere soil and on the root surface of grapevines.

  7. Strain-level diversity of secondary metabolism in the biocontrol species Aneurinibacillus migulanus.

    Science.gov (United States)

    Alenezi, Faizah N; Rekik, Imen; Bełka, Marta; Ibrahim, Abrar F; Luptakova, Lenka; Jaspars, Marcel; Woodward, Steve; Belbahri, Lassaad

    2016-01-01

    Aneurinibacillus migulanus strains Nagano and NCTC 7096 show potential in biocontrol against fungal and fungus-like plant pathogens, including a wide range of Ascomycota, Basidiomycota, and Oomycetes. Differences in terms of the range of pathogens that each strain inhibits, however, suggested that production of a single antibiotic cyclic peptide, gramicidin S (GS), by the two strains, is not the sole mechanism of inhibition. The availability of four sequenced genomes of Aneurinibacillus prompted us to apply genome mining techniques to identify the bioactive potential of A. migulanus and to provide insights into the secondary metabolite arsenal of the genus Aneurinibacillus. Up to eleven secondary metabolite biosynthetic gene clusters were present in the three Aneurinibacillus species. Biosynthetic gene clusters specifying bacteriocins, microcins, non-ribosomal peptides, polyketides, terpenes, phosphonates, lasso peptides and linaridins were identified. Chitinolytic potential and iron metabolism regulation were also investigated. With increasing numbers of biocontrol bacterial genomes being sequenced and mined, the use of approaches similar to those described in this paper will lead to an increase in the numbers of environmentally friendly natural products available to use against plant diseases. PMID:26686620

  8. Cell wall degrading isoenzyme profiles of Trichoderma biocontrol strains show correlation with rDNA species

    Institute of Scientific and Technical Information of China (English)

    Sanz L; Hermosa M R; González F J; Monte E

    2004-01-01

    @@ Species of the fungus Trichoderma, a genus of Hyphomycetes, are ubiquitous in the environment, but especially in soil. They have been used in a wide range of commercial applications including the production of hydrolases and in the biological control of plant diseases. A fundamental part of the Trichoderma antifungal system consists of a series of genes coding for a surprising variety of extracellular cell wall degrading enzymes (CWDE).Characterisation and identification of strains at the species level is the first step in utilizing the full potential of fungi in specific applications. One aim when isolating Trichoderma strains is to identify those which can be used in new agricultural and industrial applications. In the past it was not uncommon that biocontrol strains were defined as T. harzianum Rifai, due to the limited classification system of the genus Trichoderma. In recent years, several PCR-based molecular techniques have been used to detect and discriminate among microorganisms. Sequence analysis of the ITS regions of the ribosomal DNA and gene fragments as those corresponding to tef1 gene have been helpful in the neotypification, description and characterization of species in the genus Trichoderna.Another useful method for the identification of Trichoderma strains is the randomly amplified polymorphic DNA (RAPD) technique.Isozyme polymorphisms evaluation of five putative extracellular lytic enzymes loci (β-1,3-glucanase, β-1,6-glucanase, cellulase, chitinase and protease antivities) were carried out using representative strains of defined molecular groups. CWDE groupings obtained from biocontrol strains are discussed in relation to their phylogenetic location and antifungal activities.Compiling morphological, biochemical and sequence information data into a common database would provide a useful resource that could be used to accurately name new haplotypes identified in the future and correctly place them within the genus Trichoderma.

  9. Biocontrol of Aspergillus flavus on peanut kernels by use of a strain of marine Bacillus megaterium.

    Science.gov (United States)

    Kong, Qing; Shan, Shihua; Liu, Qizheng; Wang, Xiudan; Yu, Fangtang

    2010-04-30

    A strain of marine Bacillus megaterium isolated from the Yellow Sea of East China was evaluated for its activity in reducing postharvest decay of peanut kernels caused by Aspergillus flavus in in vitro and in vivo tests. The results showed that the concentrations of antagonist had a significant effect on biocontrol effectiveness in vivo: when the concentration of the washed bacteria cell suspension was used at 1x10(9)CFU/ml, the percentage rate of rot of peanut kernels was 31.67%+/-2.89%, which was markedly lower than that treated with water (the control) after 7days of incubation at 28 degrees C. The results also showed that unwashed cell culture of B. megaterium was as effective as the washed cell suspension, and better biocontrol was obtained when longer incubation time of B. megaterium was applied. When the incubation time of B. megaterium was 60-h, the rate of decay declined to 41.67%+/-2.89%. Furthermore, relative to the expression of 18S rRNA, the mRNA abundances of aflR gene and aflS gene in the experiment group were 0.28+/-0.03 and 0.024+/-0.005 respectively, indicating that this strain of B. megaterium could significantly reduce the biosynthesis of aflatoxins and expression of aflR gene and aflS gene (pagainst postharvest fungal disease caused by A. flavus. PMID:20156660

  10. Systematics of the Trichoderma harzianum species complex and the re-identification of commercial biocontrol strains.

    Science.gov (United States)

    Chaverri, Priscila; Branco-Rocha, Fabiano; Jaklitsch, Walter; Gazis, Romina; Degenkolb, Thomas; Samuels, Gary J

    2015-01-01

    Trichoderma harzianum is known as a cosmopolitan, ubiquitous species associated with a wide variety of substrates. It is possibly the most commonly used name in agricultural applications involving Trichoderma, including biological control of plant diseases. While various studies have suggested that T. harzianum is a species complex, only a few cryptic species are named. In the present study the taxonomy of the T. harzianum species complex is revised to include at least 14 species. Previously named species included in the complex are T. guizhouense, T. harzianum, and T. inhamatum. Two new combinations are proposed, T. lentiforme and T. lixii. Nine species are described as new, T. afarasin, T. afroharzianum, T. atrobrunneum, T. camerunense, T. endophyticum, T. neotropicale, T. pyramidale, T. rifaii and T. simmonsii. We isolated Trichoderma cultures from four commercial biocontrol products reported to contain T. harzianum. None of the biocontrol strains were identified as T. harzianum s. str. In addition, the widely applied culture 'T. harzianum T22' was determined to be T. afroharzianum. Some species in the T. harzianum complex appear to be exclusively endophytic, while others were only isolated from soil. Sexual states are rare. Descriptions and illustrations are provided. A secondary barcode, nuc translation elongation factor 1-α (TEF1) is needed to identify species in this complex.

  11. Complete Genome Sequence of Pseudomonas fluorescens LBUM636, a Strain with Biocontrol Capabilities against Late Blight of Potato.

    Science.gov (United States)

    Morrison, Christopher K; Novinscak, Amy; Gadkar, Vijay J; Joly, David L; Filion, Martin

    2016-01-01

    Herein provided is the full-genome sequence of Pseudomonas fluorescens LBUM636. This strain is a plant growth-promoting rhizobacterium (PGPR) which produces phenazine-1-carboxylic acid, an antibiotic involved in the biocontrol of numerous plant pathogens, including late blight of potato caused by the plant pathogen Phytophthora infestans. PMID:27231373

  12. Biocontrol of geosmin-producing Streptomyces spp. by two Bacillus strains from Chinese liquor.

    Science.gov (United States)

    Zhi, Yan; Wu, Qun; Du, Hai; Xu, Yan

    2016-08-16

    Streptomyces spp. producing geosmin have been regarded as the most frequent and serious microbial contamination causing earthy off-flavor in Chinese liquor. It is therefore necessary to control the Streptomyces community during liquor fermentation. Biological control, using the native microbiota present in liquor making, appears to be a better solution than chemical methods. The objective of this study was to isolate native microbiota antagonistic toward Streptomyces spp. and then to evaluate the possible action mode of the antagonists. Fourteen Bacillus strains isolated from different Daqu (the fermentation starter) showed antagonistic activity against Streptomyces sampsonii, which is one of the dominant geosmin producers. Bacillus subtilis 2-16 and Bacillus amyloliquefaciens 1-45 from Maotai Daqu significantly inhibited the growth of S. sampsonii by 57.8% and 84.3% respectively, and effectively prevented the geosmin production in the simulated fermentation experiments (inoculation ratio 1:1). To probe the biocontrol mode, the ability of strain 2-16 and 1-45 to produce antimicrobial metabolites and to reduce geosmin in the fermentation system was investigated. Antimicrobial substances were identified as lipopeptides by ultra-performance liquid chromatography tandem electrospray ionization/quadrupole-time-of-flight mass spectrometry (UPLC-ESI/Q-TOF MS) and in vitro antibiotic assay. In addition, strains 2-16 and 1-45 were able to remove 45% and 15% of the geosmin respectively in the simulated solid-state fermentation. This study highlighted the potential of biocontrol, and how the use of native Bacillus species in Daqu could provide an eco-friendly method to prevent growth of Streptomyces spp. and geosmin contamination in Chinese liquor fermentation.

  13. In Vitro Screening for Abiotic Stress Tolerance in Potent Biocontrol and Plant Growth Promoting Strains of Pseudomonas and Bacillus spp.

    OpenAIRE

    G. Praveen Kumar; Mir Hassan Ahmed, S. K.; Suseelendra Desai; Leo Daniel Amalraj, E.; Abdul Rasul

    2014-01-01

    Plant growth promoting rhizobacteria (PGPR) has been identified as a group of microbes that are used for plant growth enhancement and biocontrol for management of plant diseases. The inconsistency in performance of these bacteria from laboratory to field conditions is compounded due to the prevailing abiotic stresses in the field. Therefore, selection of bacterial strains with tolerance to abiotic stresses would benefit the end-user by successful establishment of the strain for showing desire...

  14. Characterization of antagonistic-potential of two Bacillus strains and their biocontrol activity against Rhizoctonia solani in tomato.

    Science.gov (United States)

    Solanki, Manoj Kumar; Singh, Rajesh Kumar; Srivastava, Supriya; Kumar, Sudheer; Kashyap, Prem Lal; Srivastava, Alok K

    2015-01-01

    To investigate the biocontrol mechanism of two antagonistic Bacillus strains (Bacillus subtilis MB14 and Bacillus amyloliquefaciens MB101), three in vitro antagonism assays were screened and the results were concluded that both strains inhibited Rhizoctonia solani growth in a similar manner by dual culture assay, but the maximum percent of inhibition only resulted with MB101 by volatile and diffusible metabolite assays. Moreover, cell free supernatant (CFS) of MB101 also showed significant (p > 0.05) growth inhibition as compared to MB14, when 10 and 20% CFS mix with the growth medium of R. solani. After in vitro-validation, both strains were evaluated under greenhouse and the results concluded that strain MB101 had significant biocontrol potential as compared to MB14. Strain MB101 was enhanced the plant height, biomass and chlorophyll content of tomato plant through a higher degree of root colonization. In field trials, strain MB101 showed higher lessening in root rot symptoms with significant fruit yield as compare to strain MB14 and infected control. Next to the field study, the presence of four antibiotic genes (srfAA, fenD, ituC, and bmyB) also concluded the antifungal nature of both Bacillus strains. Phylogenetic analysis of protein sequences revealed a close relatedness of three genes (srfAA, fenD, and ituC) with earlier reported sequences of B. subtilis and B. amyloliquefaciens. However, bmyB showed heterogeneity in among both strains (MB14 and MB101) and it may be concluded that higher degree of antagonism, root colonization and different antibiotic producing genes may play an important role in biocontrol mechanism of strain MB101. PMID:24277414

  15. Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1.

    Directory of Open Access Journals (Sweden)

    Shujian Zhang

    Full Text Available Xylella fastidiosa (X. fastidiosa infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703 and a serine protease (PD0956; two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928, and at least one relatively short, hemagglutinin-like protein (PD0986. Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3 overexpressing PD1703 exhibited a hypersensitive response (HR in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot and a PD0986 (hemagglutinin were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry.

  16. Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1.

    Science.gov (United States)

    Zhang, Shujian; Chakrabarty, Pranjib K; Fleites, Laura A; Rayside, Patricia A; Hopkins, Donald L; Gabriel, Dean W

    2015-01-01

    Xylella fastidiosa (X. fastidiosa) infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD) of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703) and a serine protease (PD0956); two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928), and at least one relatively short, hemagglutinin-like protein (PD0986). Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3) overexpressing PD1703 exhibited a hypersensitive response (HR) in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot) and a PD0986 (hemagglutinin) were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry. PMID:26218423

  17. Draft Genome Sequences of Pseudomonas fluorescens Strains SF39a and SF4c, Potential Plant Growth Promotion and Biocontrol Agents.

    Science.gov (United States)

    Ly, Lindsey K; Underwood, Grace E; McCully, Lucy M; Bitzer, Adam S; Godino, Agustina; Bucci, Vanni; Brigham, Christopher J; Príncipe, Analía; Fischer, Sonia E; Silby, Mark W

    2015-03-26

    Pseudomonas fluorescens SF4c and SF39a, strains isolated from wheat rhizosphere, have potential applications in plant growth promotion and biocontrol of fungal diseases of crop plants. We report the draft genome sequences of SF4c and SF39a with estimated sizes of 6.5 Mb and 5.9 Mb, respectively.

  18. Draft Genome Sequence of Cryptococcus flavescens Strain OH182.9_3C, a Biocontrol Agent against Fusarium Head Blight of Wheat

    OpenAIRE

    Rong, Xiaoqing; McSpadden Gardener, Brian B.

    2013-01-01

    Cryptococcus flavescens strain OH182.9_3C (3C) is a novel biopesticidal agent that can be used to control fusarium head blight of wheat. Here we present the draft genome sequence for 3C, the first for the species C. flavescens. Additionally, several genes that may contribute to the biocontrol activities of 3C were identified in silico.

  19. Colonization pattern of the biocontrol strain Pseudomonas chlororaphis MA 342 on barley seeds visualized by using green fluorescent protein.

    Science.gov (United States)

    Tombolini, R; van der Gaag, D J; Gerhardson, B; Jansson, J K

    1999-08-01

    Pseudomonas chlororaphis MA 342 is a potent biocontrol agent that can be used against several seed-borne diseases of cereal crops, including net blotch of barley caused by the fungus Drechslera teres. In this study, strain MA 342 was tagged with the gfp gene (encoding the green fluorescent protein) in order to study the fate of cells after seed inoculation. The gfp-tagged strain, MA 342G2, had the same biocontrol efficacy as the wild type when it was applied at high cell concentrations to seeds but was less effective at lower cell concentrations. By comparing cell counts determined by microscopy to the number of CFU, we found that the number of culturable cells was significantly lower than the total number of bacteria on seeds which were inoculated and dried for 20 h. Confocal microscopy and epifluorescence stereomicroscopy were used to determine the pattern of MA 342G2 colonization and cell aggregation on barley seeds. Immediately after inoculation of seeds, bacteria were found mainly under the seed glume, and there was no particular aggregation pattern. However, after the seeds were sown, irregularly distributed areas of bacterial aggregation were found, which reflected epiphytic colonization of glume cells. There was a trend towards bacterial aggregation near the embryo but never within the embryo. Bacterial aggregates were regularly found in the groove of each seed formed by the base of the coleoptile and the scutellum. Based on these results, we suggest that MA 342 colocalizes with the pathogen D. teres, which facilitates the action of the fungistatic compound(s) produced by this strain.

  20. Screening for Pseudomonas and Bacillus antagonistic rhizobacteria strains for the biocontrol of Fusarium wilt of chickpea

    Directory of Open Access Journals (Sweden)

    Hannane Abed

    2016-07-01

    Full Text Available The aim of this work is to study the ability of several isolates belonging to Rhizobacteria (Pseudomonas and Bacillus collected from several chickpea growing areas in Algeria, to control the mycelium growth of Fusarium oxysporum f. sp. ciceris. Interesting isolates were characterized for their morphological characteristics, physiological and biochemical activities as potential bio-control agent. Fungal inhibition tests were performed using plate assay and each isolate were tested for the production of protease, cyanide hydrogen, indole acetic acid, antifungal volatile and extracellular compound. According to API 50 CH, we are able to identify six Bacillus species (B. subtilis, B. circulans, B. lentus, B. aneurinilyticus, B. firmus, B. licheniformis; and with API 20NE test we have identified three Pseudomonas species (P. aeruginosa, P. luteola, P. fluorescens. The ability of bacterial isolates was varied in production of Protease, Gelatinase, Amylase, Cellulase, Acid Indole acetic, Lipase, Catalase and Cyanid Hydrogen. This is traduced in different rate of inhibition growth due to various extracellular compounds, where B61 (Bacillus aneurinilyticus and P39 (Pseudomonas luteola and P70 (Pseudomonas fluorescens were the most efficient with 77 and 55.5% respectively, while B39 (Bacillus firmus and P41 (Pseudomonas luteola were the most efficient by volatile compounds with 70.5 and 77.5% respectively. Our results indicate that these bacteria isolates can be used in the biocontrol of Fusarium oxysporum f. sp. ciceris.

  1. Use of Hypocrea jecorina (anamorph Trichoderma reesei) as a model system for Trichoderma biocontrol of Pythium blight identifies new targets for genetic strain improvement

    Institute of Scientific and Technical Information of China (English)

    Seidl V; Schmoll M; Scherm B; Balmas V; Seiboth B; Migheli Q; Kubicek C P

    2004-01-01

    @@ Biocontrol by Trichoderma has been studied mainly with selected isolates of T. harzianum, T.atroviride and T. asperellum, which are members of sections Pachybasium and Trichoderma. In contrast, species from section Longibrachiatum have only rarely been studied. On the other hand, one taxon from this section-Hypocrea jecorina (anamorph: Trichoderma reesei)-has been widely used for the production of cellulolytic and hemicellulolytic enzymes and recombinant proteins. As far as Trichoderma is concerned, molecular genetic methods and tools are most advanced in H. jecorina,and its genome has recently been fully sequenced, thus making this taxon a model organism for the genus. Here we will demonstrate that H. jecorina is able to antagonize plant pathogenic fungi in plate confrontation tests, and can protect tomato and cucumber plants against Pythium ultimum blight.Using this as a model case, we made use of available H. jecorina mutants to investigate (a) whether carbon catabolite repression via the Cre1-regulator protein has an impact on biocontrol, and (b)whether cellulase gene expression is necessary for biocontrol of P. ultimum. In the first case, plate confrontation tests and in planta experiments yielded opposite results, i.e. while a Cre1 mutant was more active in antagonization of fungi on plates, the survival rates of P. ultimum-inoculated cucumber plants was lower than with the H. jecorina wild-type strain. Mutants of H. jecorina,unable to form cellulases, were still able to antagonize fungi on plates and provided similar protection of tomatos against P. ultimum as the wild type, indicating that the pronounced biocontrol ability of H.jecorina against fungi with cellulose-containing cell-walls is not due to its high cellulolytic activity. A strain disrupted in the light-modulator gene envoy (Schmoll et al., ms submitted) exhibited in planta biocontrol activity strongly exceeding that of the wild-type strain, thereby providing a first link between Trichoderma

  2. Complete genome sequence of Pseudomonas fluorescens strain PICF7, an indigenous root endophyte from olive (Olea europaea L.) and effective biocontrol agent against Verticillium dahliae.

    Science.gov (United States)

    Martínez-García, Pedro Manuel; Ruano-Rosa, David; Schilirò, Elisabetta; Prieto, Pilar; Ramos, Cayo; Rodríguez-Palenzuela, Pablo; Mercado-Blanco, Jesús

    2015-01-01

    Pseudomonas fluorescens strain PICF7 is a native endophyte of olive roots. Previous studies have shown this motile, Gram-negative, non-sporulating bacterium is an effective biocontrol agent against the soil-borne fungus Verticillium dahliae, the causal agent of one of the most devastating diseases for olive (Olea europaea L.) cultivation. Here, we announce and describe the complete genome sequence of Pseudomonas fluorescens strain PICF7 consisting of a circular chromosome of 6,136,735 bp that encodes 5,567 protein-coding genes and 88 RNA-only encoding genes. Genome analysis revealed genes predicting factors such as secretion systems, siderophores, detoxifying compounds or volatile components. Further analysis of the genome sequence of PICF7 will help in gaining insights into biocontrol and endophytism.

  3. Biocontrol ability and action mechanism of food-isolated yeast strains against Botrytis cinerea causing post-harvest bunch rot of table grape.

    Science.gov (United States)

    Parafati, Lucia; Vitale, Alessandro; Restuccia, Cristina; Cirvilleri, Gabriella

    2015-05-01

    Strains belonging to the species Saccharomyces cerevisiae, Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans, isolated from different food sources, were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic mold Botrytis cinerea. All yeast strains demonstrated antifungal activity at different levels depending on species and medium. Killer strains of W. anomalus and S. cerevisiae showed the highest biocontrol in vitro activity, as demonstrated by largest inhibition halos. The competition for iron and the ability to form biofilm and to colonize fruit wounds were hypothesized as the main action mechanisms for M. pulcherrima. The production of hydrolytic enzymes and the ability to colonize the wounds were the most important mechanisms for biocontrol activity in A. pullulans and W. anomalus, which also showed high ability to form biofilm. The production of volatile organic compounds (VOCs) with in vitro and in vivo inhibitory effect on pathogen growth was observed for the species W. anomalus, S. cerevisiae and M. pulcherrima. Our study clearly indicates that multiple modes of action may explain as M. pulcherrima provide excellent control of postharvest botrytis bunch rot of grape. PMID:25583341

  4. Production of trichodiene by Trichoderma harzianum alters the perception of this biocontrol strain by plants and antagonized fungi.

    Science.gov (United States)

    Malmierca, Mónica G; McCormick, Susan P; Cardoza, Rosa E; Alexander, Nancy J; Monte, Enrique; Gutiérrez, Santiago

    2015-08-01

    Trichothecenes are phytotoxic sesquiterpenic mycotoxins that can act as virulence factors in plant diseases. Harzianum A (HA) is a non-phytotoxic trichothecene produced by Trichoderma arundinaceum. The first step in HA biosynthesis is the conversion of farnesyl diphosphate to trichodiene (TD), a volatile organic compound (VOC), catalysed by a sesquiterpene synthase encoded by the tri5 gene. Expression of tri5 in the biocontrol strain Trichoderma harzianum CECT 2413 resulted in production of TD in parallel with a reduction of ergosterol biosynthesis and an unexpected increase in the level of squalene. Transformants expressing tri5 displayed low chitinase activity and induced expression of Botrytis cinerea BOT genes, although their total antagonistic potential against phytopathogenic fungi was not reduced. VOCs released by the tri5-transformant induced expression of tomato defence genes related to salicylic acid (SA), and TD itself strongly induced the expression of SA-responsive genes and reduced the development of lateral roots. Together, these results suggest that TD acts as a signalling VOC in the interactions of Trichoderma with plants and other microorganisms by modulating the perception of this fungus to a given environment. Moreover, the TD ability to induce systemic defences indicates that complex trichothecene structures may not be necessary for inducing such responses.

  5. Production of trichodiene by Trichoderma harzianum alters the perception of this biocontrol strain by plants and antagonized fungi.

    Science.gov (United States)

    Malmierca, Mónica G; McCormick, Susan P; Cardoza, Rosa E; Alexander, Nancy J; Monte, Enrique; Gutiérrez, Santiago

    2015-08-01

    Trichothecenes are phytotoxic sesquiterpenic mycotoxins that can act as virulence factors in plant diseases. Harzianum A (HA) is a non-phytotoxic trichothecene produced by Trichoderma arundinaceum. The first step in HA biosynthesis is the conversion of farnesyl diphosphate to trichodiene (TD), a volatile organic compound (VOC), catalysed by a sesquiterpene synthase encoded by the tri5 gene. Expression of tri5 in the biocontrol strain Trichoderma harzianum CECT 2413 resulted in production of TD in parallel with a reduction of ergosterol biosynthesis and an unexpected increase in the level of squalene. Transformants expressing tri5 displayed low chitinase activity and induced expression of Botrytis cinerea BOT genes, although their total antagonistic potential against phytopathogenic fungi was not reduced. VOCs released by the tri5-transformant induced expression of tomato defence genes related to salicylic acid (SA), and TD itself strongly induced the expression of SA-responsive genes and reduced the development of lateral roots. Together, these results suggest that TD acts as a signalling VOC in the interactions of Trichoderma with plants and other microorganisms by modulating the perception of this fungus to a given environment. Moreover, the TD ability to induce systemic defences indicates that complex trichothecene structures may not be necessary for inducing such responses. PMID:24813508

  6. Novel Trichoderma polysporum Strain for the Biocontrol of Pseudogymnoascus destructans, the Fungal Etiologic Agent of Bat White Nose Syndrome.

    Directory of Open Access Journals (Sweden)

    Tao Zhang

    Full Text Available White-nose syndrome (WNS, an emerging disease of hibernating bats, has rapidly spread across eastern North America killing millions of bats. Pseudogymnoascus destructans (Pd, the sole etiologic agent of WNS, is widespread and persistent in bat hibernacula. Control of Pd in the affected sites is urgently needed to break the transmission cycle while minimizing any adverse impact on the native organisms. We isolated a novel strain of Trichoderma polysporum (Tp from one of the caves at the epicenter of WNS zoonotic. Detailed experimental studies revealed: (1 Tp WPM 39143 was highly adapted to grow at temperatures simulating the cave environment (6°C-15°C, (2 Tp WPM 39143 restricted Pd colony growth in dual culture challenges, (3 Tp WPM 39143 caused four logs reduction of Pd colony forming units and genome copies in autoclaved soil samples from one of the WNS affected caves, (4 Tp WPM 39143 extract showed specific fungicidal activity against Pd in disk diffusion assay, but not against closely related fungus P. pannorum (Pp, (5 Tp WPM 39143 extract retained inhibitory activity after exposure to high temperatures, light and proteinase K, and (6 Inhibitory metabolites in Tp WPM 39143 extract comprised of water-soluble, high polarity compounds. These results suggest that Tp WPM 39143 is a promising candidate for further evaluation as a biocontrol agent of Pd in WNS affected sites.

  7. Molecular Identification Of Trichoderma Strains Collected To Develop Plant Growth-Promoting And Biocontrol Agents

    Directory of Open Access Journals (Sweden)

    Oskiera Michał

    2015-06-01

    Full Text Available Trichoderma strains that are beneficial to both the growth and health of plants can be used as plant growth-promoting fungi (PGPF or biological control agents (BCA in agricultural and horticultural practices. In order to select PGPF or BCA strains, their biological properties and taxonomy must be carefully studied. In this study, 104 strains of Trichoderma collected at geographically different locations in Poland for selection as PGPF or BCA were identified by DNA barcoding, based on the sequences of internal transcribed spacers 1 and 2 (ITS1 and 2 of the ribosomal RNA gene cluster and on the sequences of translation elongation factor 1 alpha (tef1, chitinase 18-5 (chi18-5, and RNA polymerase II subunit (rpb2 gene fragments. Most of the strains were classified as: T. atroviride (38%, T. harzianum (21%, T. lentiforme (9%, T. virens (9%, and T. simmonsii (6%. Single strains belonging to T. atrobrunneum, T. citrinoviride, T. crassum, T. gamsii, T. hamatum, T. spirale, T. tomentosum, and T. viridescens were identified. Three strains that are potentially pathogenic to cultivated mushrooms belonging to T. pleuroticola and T. aggressivum f. europaeum were also identified. Four strains: TRS4, TRS29, TRS33, and TRS73 were classified to Trichoderma spp. and molecular identification was inconclusive at the species level. Phylogeny analysis showed that three of these strains TRS4, TRS29, and TRS33 belong to Trichoderma species that is not yet taxonomically established and strain TRS73 belongs to the T. harzianum complex, however, the species could not be identified with certainty.

  8. Requirement of Simultaneous Assessment of Crystal- and Supernatant-Related Entomotoxic Activities of Bacillus thuringiensis Strains for Biocontrol-Product Development

    Directory of Open Access Journals (Sweden)

    Ronaldo Costa Argôlo-Filho

    2014-05-01

    Full Text Available Bioinsecticides with lower concentrations of endospores/crystals and without loss of efficiency are economically advantageous for pest biocontrol. In addition to Cry proteins, other Bacillus thuringiensis (Bt toxins in culture supernatants (SN have biocontrol potential (e.g., Vip3A, Cry1I, Sip1, whereas others are unwanted (β-exotoxins, as they display widespread toxicity across taxa. A strain simultaneously providing distinct toxin activities in crystals and SN would be desirable for bioinsecticides development; however, strains secreting β-exotoxins should be discarded, independently of other useful entomotoxins. Entomotoxicity of crystals and SN from a Brazilian Bt tolworthi strain (Btt01 was tested against Spodoptera frugiperda to assess the potential for biocontrol-product development based on more than one type of toxin/activity. Tests showed that 107 endospores mL−1 caused >80% of larvae mortality, suggesting Btt01 may be used in similar concentrations as those of other Bt-based biopesticides. When it was applied to cornfields, a significant 60% reduction of larvae infestation was observed. However, bioassays with Btt01 SN revealed a thermostable toxic activity. Physicochemical characterization strongly suggests the presence of unwanted β-exotoxins, with isolate-specific temporal variation in its secretion. Knowledge of the temporal pattern of secretion/activity in culture for all forms of toxins produced by a single strain is required to both detect useful activities and avoid the potential lack of identification of undesirable toxins. These findings are discussed in the contexts of commercial Bt product development, advantages of multiple-activity strains, and care and handling recommended for large-scale fermentation systems.

  9. Evaluation of Pseudomonas syringae Strain ESC11 for Biocontrol of Crown Rot and Anthracnose of Banana

    Science.gov (United States)

    Pseudomonas syringae strain ESC11, and 250 'g/ml each of thiabendazole (TBZ) and imazalil reduced crown rot of banana caused by a Fusarium sp. by 0-88% and 73-88%, respectively, in laboratory experiments. ESC11 alone did not significantly reduce rot, mold, or anthracnose in most field trials. TBZ an...

  10. Genome Sequencing of a Mung Bean Plant Growth Promoting Strain of P. aeruginosa with Biocontrol Ability

    OpenAIRE

    Devaraj Illakkiam; Manoharan Shankar; Paramasivan Ponraj; Jeyaprakash Rajendhran; Paramasamy Gunasekaran

    2014-01-01

    Pseudomonas aeruginosa PGPR2 is a mung bean rhizosphere strain that produces secondary metabolites and hydrolytic enzymes contributing to excellent antifungal activity against Macrophomina phaseolina, one of the prevalent fungal pathogens of mung bean. Genome sequencing was performed using the Ion Torrent Personal Genome Machine generating 1,354,732 reads (6,772,433 sequenced bases) achieving ~25-fold coverage of the genome. Reference genome assembly using MIRA 3.4.0 yielded 198 contigs. The ...

  11. Analysis of Clonostachys rosea-induced resistance to tomato gray mold disease in tomato leaves.

    Directory of Open Access Journals (Sweden)

    Liana Dalcantara Ongouya Mouekouba

    Full Text Available Tomato gray mold disease, caused by Botrytis cinerea, is a serious disease in tomato. Clonostachys rosea is an antagonistic microorganism to B. cinerea. To investigate the induced resistance mechanism of C. rosea, we examined the effects of these microorganisms on tomato leaves, along with changes in the activities of three defense enzymes (PAL, PPO, GST, second messengers (NO, H2O2, O2(- and phytohormones (IAA, ABA, GA3, ZT, MeJA, SA and C2H4. Compared to the control, all treatments induced higher levels of PAL, PPO and GST activity in tomato leaves and increased NO, SA and GA3 levels. The expression of WRKY and MAPK, two important transcription factors in plant disease resistance, was upregulated in C. rosea- and C. rosea plus B. cinerea-treated samples. Two-dimensional gel electrophoresis analysis showed that two abundant proteins were present in the C. rosea plus B. cinerea-treated samples but not in the other samples. These proteins were determined (by mass spectrum analysis to be LEXYL2 (β-xylosidase and ATP synthase CF1 alpha subunit. Therefore, C. rosea plus B. cinerea treatment induces gray mold resistance in tomato. This study provides a basis for elucidating the mechanism of C. rosea as a biocontrol agent.

  12. Genome Sequencing of a Mung Bean Plant Growth Promoting Strain of P. aeruginosa with Biocontrol Ability

    Directory of Open Access Journals (Sweden)

    Devaraj Illakkiam

    2014-01-01

    Full Text Available Pseudomonas aeruginosa PGPR2 is a mung bean rhizosphere strain that produces secondary metabolites and hydrolytic enzymes contributing to excellent antifungal activity against Macrophomina phaseolina, one of the prevalent fungal pathogens of mung bean. Genome sequencing was performed using the Ion Torrent Personal Genome Machine generating 1,354,732 reads (6,772,433 sequenced bases achieving ~25-fold coverage of the genome. Reference genome assembly using MIRA 3.4.0 yielded 198 contigs. The draft genome of PGPR2 encoded 6803 open reading frames, of which 5314 were genes with predicted functions, 1489 were genes of known functions, and 80 were RNA-coding genes. Strain specific and core genes of P. aeruginosa PGPR2 that are relevant to rhizospheric habitat were identified by pangenome analysis. Genes involved in plant growth promoting function such as synthesis of ACC deaminase, indole-3-acetic acid, trehalose, mineral scavenging siderophores, hydrogen cyanide, chitinases, acyl homoserine lactones, acetoin, 2,3-butanediol, and phytases were identified. In addition, niche-specific genes such as phosphate solubilising 3-phytase, adhesins, pathway-specific transcriptional regulators, a diguanylate cyclase involved in cellulose synthesis, a receptor for ferrienterochelin, a DEAD/DEAH-box helicase involved in stress tolerance, chemotaxis/motility determinants, an HtpX protease, and enzymes involved in the production of a chromanone derivative with potent antifungal activity were identified.

  13. Bacterial Feeders, the Nematode Caenorhabditis elegans and the Flagellate Cercomonas longicauda, have different Effects on Outcome of Competition among the Pseudomonas Biocontrol Strains CHA0 and DSS73

    DEFF Research Database (Denmark)

    Pedersen, Annette; Nybroe, Ole; Winding, Anne;

    2009-01-01

    selective feeding flagellate Cercomonas longicauda versus the non-selective feeding nematode Caenorhabditis elegans) influence the abundance of two bacteria that compete for resources in simple model communities. Microcosms consisted of either one gfp-tagged bacterial strain (Pseudomonas fluorescens DSM......50090 or one of two biocontrol strains P. fluorescens CHA0 or Pseudomonas sp. DSS73) or combinations of two bacterial strains. DSM50090 is a suitable food bacterium, DSS73 is of intermediate food quality, and CHA0 is inedible to the bacterial feeders. Bacterial and protozoan cell numbers were measured......How bacterial feeding fauna affects colonization and survival of bacteria in soil is not well understood, which constrains the applicability of bacterial inoculants in agriculture. This study aimed to unravel how food quality of bacteria and bacterial feeders with different feeding habits (the...

  14. A Perilipin Gene from Clonostachys rosea f. Catenulata HL-1-1 Is Related to Sclerotial Parasitism

    Directory of Open Access Journals (Sweden)

    Zhan-Bin Sun

    2015-03-01

    Full Text Available Clonostachys rosea f. catenulata is a promising biocontrol agent against many fungal plant pathogens. To identify mycoparasitism-related genes from C. rosea f. catenulata, a suppression subtractive hybridization (SSH cDNA library of C. rosea f. catenulata HL-1-1 that parasitizes the sclerotia of S. sclerotiorum was constructed. 502 clones were sequenced randomly, and thereby 472 expressed sequence tags (ESTs were identified. Forty-three unigenes were annotated and exhibited similarity to a wide diversity of genes. Quantitative real -time PCR showed that a perilipin-like protein encoding gene, Per3, was up-regulated by 6.6-fold over the control at 96 h under the induction of sclerotia. The full-length sequence of Per3 was obtained via 5' and 3' rapid identification of cDNA ends. Overexpression of Per3 in HL-1-1 significantly enhanced the parasitic ability on sclerotia. The results indicated that Per3 might be involved in the mycoparasitism of C. rosea f. catenulata HL-1-1. This is the first report of a perilipin as a potential biocontrol gene in mycoparasites. The study provides usefu l insights into the interaction between C. rosea f. catenulata and fungal plant pathogens.

  15. A phenazine-1-carboxylic acid producing polyextremophilic Pseudomonas chlororaphis (MCC2693) strain, isolated from mountain ecosystem, possesses biocontrol and plant growth promotion abilities.

    Science.gov (United States)

    Jain, Rahul; Pandey, Anita

    2016-09-01

    The genus Pseudomonas is known to comprise a huge diversity of species with the ability to thrive in different habitats, including those considered as extreme environments. In the present study, a psychrotolerant, wide pH tolerant and halotolerant strain of Pseudomonas chlororaphis GBPI_507 (MCC2693), isolated from the wheat rhizosphere growing in a mountain location in Indian Himalayan Region (IHR), has been investigated for its antimicrobial potential with particular reference to phenazine production and plant growth promoting traits. GBPI_507 showed phenazine production at the temperatures ranged from 14 to 25°C. The benzene extracted compound identified as phenazine-1-carboxylic acid (PCA) through GC-MS exhibited antimicrobial properties against Gram positive bacteria and actinomycetes. The inhibition of phytopathogens in diffusible biocontrol assays was recorded in an order: Alternaria alternata>Phytophthora sp.>Fusarium solani>F. oxysporum. In volatile metabolite assays, all the pathogens, except Phytophthora sp. produced distorted colonies, characterized by restricted sporulation. The isolate also possessed other growth promoting and biocontrol traits including phosphate solubilization and production of siderophores, HCN, ammonia, and lytic enzymes (lipase and protease). Molecular studies confirmed production of PCA by the bacterium GBPI_507 through presence of phzCD and phzE genes in its genome. The polyextremophilic bacterial strain possesses various important characters to consider it as a potential agent for field applications, especially in mountain ecosystem, for sustainable and eco-friendly crop production. PMID:27394000

  16. Fusarium oxysporum strains as biocontrol agents against Fusarium wilt: effects on soil microbial biomass and activity Linhagens de Fusarium oxysporum como agentes de biocontrole da murcha-de-Fusarium: efeitos na biomassa e atividade microbiana do solo

    Directory of Open Access Journals (Sweden)

    Raquel Ghini

    2000-01-01

    Full Text Available Before planning the large-scale use of nonpathogenic strains of Fusarium oxysporum as biocontrol agents of Fusarium wilt, their behaviour and potential impact on soil ecosystems should be carefully studied as part of risk assessment. The aim of this work was to evaluate the effects of antagonistic F. oxysporum strains, genetically manipulated (T26/6 or not (233/1, on soil microbial biomass and activity. The effects were evaluated, in North-western Italy, in two soils from different sites at Albenga, one natural and the other previously solarized, and in a third soil obtained from a 10-year-old poplar stand (Popolus sp., near Carignano. There were no detectable effects on ATP, fluorescein diacetate hydrolysis, and biomass P that could be attributed to the introduction of the antagonists. A transient increase of carbon dioxide evolution and biomass C was observed in response to the added inoculum. Although the results showed only some transient alterations, further studies are required to evaluate effects on specific microorganism populations.Antes do uso em larga escala de linhagens não-patogênicas de Fusarium oxysporum como agentes de biocontrole da murcha-de-Fusarium, o seu comportamento e seus impactos potenciais no solo devem ser estudados como parte da avaliação de riscos. O objetivo do presente trabalho foi avaliar os efeitos de linhagens antagonistas de F. oxysporum, geneticamente manipuladas (T26/6 ou não (233/1, na biomassa e atividade microbiana do solo. Os efeitos foram avaliados no noroeste da Itália em dois solos de Albenga, sendo um natural e outro previamente solarizado, e em um terceiro solo obtido numa plantação de 10 anos de Popolus sp., em Carignano. Não foram observados efeitos significativos na quantidade de ATP, hidrólise de diacetato de fluoresceina e biomassa P, após a introdução dos antagonistas. Um aumento transitório foi observado na evolução de dióxido de carbono e no carbono da biomassa em resposta

  17. Zearalenone detoxification by zearalenone hydrolase is important for the antagonistic ability of Clonostachys rosea against mycotoxigenic Fusarium graminearum

    DEFF Research Database (Denmark)

    Kosawang, Chatchai; Karlsson, Magnus; Vélëz, Heriberto;

    2014-01-01

    The fungus Clonostachys rosea is antagonistic against plant pathogens, including Fusarium graminearum, which produces the oestrogenic mycotoxin zearalenone (ZEA). ZEA inhibits other fungi, and C. rosea can detoxify ZEA through the enzyme zearalenone lactonohydrolase (ZHD101). As the relevance...... in vitro ability to inhibit growth of the ZEA-producing F. graminearum (strain 1104-14) compared to the wild type. In contrast, all three C. rosea strains equally inhibited growth of the F. graminearum mutant (ΔPKS4), which is impaired in ZEA-production. Furthermore, the Δzhd101 mutants failed to protect...

  18. Selection of reliable reference genes for gene expression studies in Clonostachys rosea 67-1 under sclerotial induction.

    Science.gov (United States)

    Sun, Zhan-Bin; Li, Shi-Dong; Sun, Man-Hong

    2015-07-01

    Reference genes are important to precisely quantify gene expression by real-time PCR. In order to identify stable and reliable expressed genes in mycoparasite Clonostachys rosea in different modes of nutrition, seven commonly used housekeeping genes, 18S rRNA, actin, β-tubulin, elongation factor 1, ubiquitin, ubiquitin-conjugating enzyme and glyceraldehyde-3-phosphate dehydrogenase, from the effective biocontrol isolate C. rosea 67-1 were tested for their expression under sclerotial induction and during vegetative growth on PDA medium. Analysis by three software programs showed that differences existed among the candidates. Elongation factor 1 was most stable; the M value in geNorm, SD value in Bestkeeper and stability value in Normfinder analysis were 0.405, 0.450 and 0.442, respectively, indicating that the gene elongation factor 1 could be used to normalize gene expression in C. rosea in both vegetative growth and parasitic process. By using elongation factor 1, the expression of a serine protease gene, sep, in different conditions was assessed, which was consistent with the transcriptomic data. This research provides an effective method to quantitate expression changes of target genes in C. rosea, and will assist in further investigation of parasitism-related genes of this fungus.

  19. Biocontrol of Fusarium Crown and Root Rot and Promotion of Growth of Tomato by Paenibacillus Strains Isolated from Soil

    OpenAIRE

    Xu, Sheng Jun; Kim, Byung Sup

    2014-01-01

    In this study, bacterial strains were isolated from soils from 30 locations of Samcheok, Gangwon province. Of the isolated strains, seven showed potential plant growth promoting and antagonistic activities. Based on cultural and morphological characterization, and 16S rRNA gene sequencing, these strains were identified as Paenibacillus species. All seven strains produced ammonia, cellulase, hydrocyanic acid, indole-3-acetic acid, protease, phosphatase, and siderophores. They also inhibited th...

  20. 黄曲霉毒素生物防控菌的筛选及鉴定%Screening and Identification of Aflatoxin Bio-Controlling Bacterial Strain

    Institute of Scientific and Technical Information of China (English)

    宫小明; 马荣桧; 孙军; 张艺兵; 于金玲; 丁葵英; 郭礼强

    2015-01-01

    Aflatoxin bio-controlling bacterial strain was screened in order to provide support for controlling aflatoxin. The growth,sporogenesis,and aflatoxin degradable experiments were carried out to the needed and inhibition strains screened from soil collected in peanut source land adopting Oxford-cup method. Two aflatoxin bio-controlling bacterial strains were screened and numbered as 21-1-2 and 17-3,they were characterized by authorized institution,the inhibi-tion strain 21-1-2 to be Bacillus subtilis,and inhibition strain 17-3 to be Bacillus licheniformis. They were studied on several aspects including the inhibition against the germination of Aspergillus spores,the inhibition against the growth of A. flavus,the extension of their mycelia,and the reduction of the aflatoxin production,and the degradation effects of aflatoxin. The results showed that the inhibition strains could significantly inhibit the spore germination of toxin-pro-ducing Aspergillus strains,their growth,and the extension of their mycelia,reduce the production of aflatoxin,as well as the degradation of aflatoxin.%筛选黄曲霉毒素生物防控菌,为黄曲霉毒素的生物防控提供支持。以花生原产地土壤为材料,采用牛津杯法筛选所需菌株。对筛选出的拮抗菌株进行抑制产毒曲霉菌株的生长、产孢、降解黄曲霉毒素实验。筛选出2株黄曲霉毒素生防细菌,编号21-1-2、17-3,经鉴定,拮抗菌21-1-2为枯草芽胞杆菌,拮抗菌17-3为地衣芽胞杆菌。分别对拮抗菌对曲霉孢子萌发的抑制、抑制黄曲霉的生长和菌丝延长以及减少黄曲霉毒素的产生、对黄曲霉毒素的分解作用等几个方面进行研究,结果表明,拮抗菌可以明显抑制产毒曲霉孢子的萌发、生长、菌丝的延长,减少黄曲霉毒素的产生以及分解黄曲霉毒素。

  1. Impact on Arbuscular Mycorrhiza Formation of Pseudomonas Strains Used as Inoculants for Biocontrol of Soil-Borne Fungal Plant Pathogens

    Science.gov (United States)

    Barea, J. M.; Andrade, G.; Bianciotto, V.; Dowling, D.; Lohrke, S.; Bonfante, P.; O’Gara, F.; Azcon-Aguilar, C.

    1998-01-01

    The arbuscular mycorrhizal symbiosis, a key component of agroecosystems, was assayed as a rhizosphere biosensor for evaluation of the impact of certain antifungal Pseudomonas inoculants used to control soil-borne plant pathogens. The following three Pseudomonas strains were tested: wild-type strain F113, which produces the antifungal compound 2,4-diacetylphloroglucinol (DAPG); strain F113G22, a DAPG-negative mutant of F113; and strain F113(pCU203), a DAPG overproducer. Wild-type strain F113 and mutant strain F113G22 stimulated both mycelial development from Glomus mosseae spores germinating in soil and tomato root colonization. Strain F113(pCU203) did not adversely affect G. mosseae performance. Mycelial development, but not spore germination, is sensitive to 10 μM DAPG, a concentration that might be present in the rhizosphere. The results of scanning electron and confocal microscopy demonstrated that strain F113 and its derivatives adhered to G. mosseae spores independent of the ability to produce DAPG. PMID:9603857

  2. The plant pathogenic fungus Gaeumannomyces graminis var. tritici improves bacterial growth and triggers early gene regulations in the biocontrol strain Pseudomonas fluorescens Pf29Arp.

    Science.gov (United States)

    Barret, M; Frey-Klett, P; Boutin, M; Guillerm-Erckelboudt, A-Y; Martin, F; Guillot, L; Sarniguet, A

    2009-01-01

    In soil, some antagonistic rhizobacteria contribute to reduce root diseases caused by phytopathogenic fungi. Direct modes of action of these bacteria have been largely explored; however, commensal interaction also takes place between these microorganisms and little is known about the influence of filamentous fungi on bacteria. An in vitro confrontation bioassay between the pathogenic fungus Gaeumannomyces graminis var. tritici (Ggt) and the biocontrol bacterial strain Pseudomonas fluorescens Pf29Arp was set up to analyse bacterial transcriptional changes induced by the fungal mycelium at three time-points of the interaction before cell contact and up until contact. For this, a Pf29Arp shotgun DNA microarray was constructed. Specifity of Ggt effect was assessed in comparison with one of two other filamentous fungi, Laccaria bicolor and Magnaporthe grisea. During a commensal interaction, Ggt increased the growth rate of Pf29Arp. Before contact, Ggt induced bacterial genes involved in mycelium colonization. At contact, genes encoding protein of stress response and a patatin-like protein were up-regulated. Among all the bacterial genes identified, xseB was specifically up-regulated at contact by Ggt but down-regulated by the other fungi. Data showed that the bacterium sensed the presence of the fungus early, but the main gene alteration occurred during bacterial-fungal cell contact. PMID:19121038

  3. Genome Sequence of the Endophytic Bacterium Bacillus thuringiensis Strain KB1, a Potential Biocontrol Agent against Phytopathogens

    OpenAIRE

    Jeong, Haeyoung; Jo, Sung Hee; Hong, Chi Eun; Park, Jeong Mee

    2016-01-01

    Bacillus thuringiensis is the most widely known microbial pesticide used in agricultural applications. Herein, we report a draft genome sequence of the endophytic bacterium Bacillus thuringiensis strain KB1, which exhibits antagonism against phytopathogens.

  4. Use of Lactobacillus plantarum Strains as a Bio-Control Strategy against Food-Borne Pathogenic Microorganisms.

    Science.gov (United States)

    Arena, Mattia Pia; Silvain, Amandine; Normanno, Giovanni; Grieco, Francesco; Drider, Djamel; Spano, Giuseppe; Fiocco, Daniela

    2016-01-01

    Lactobacillus plantarum is one of the most versatile species extensively used in the food industry both as microbial starters and probiotic microorganisms. Several L. plantarum strains have been shown to produce different antimicrobial compounds such as organic acids, hydrogen peroxide, diacetyl, and also bacteriocins and antimicrobial peptides, both denoted by a variable spectrum of action. In recent decades, the selection of microbial molecules and/or bacterial strains able to produce antagonistic molecules to be used as antimicrobials and preservatives has been attracting scientific interest, in order to eliminate or reduce chemical additives, because of the growing attention of consumers for healthy and natural food products. The aim of this work was to investigate the antimicrobial activity of several food-isolated L. plantarum strains, analyzed against the pathogenic bacteria Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli O157:H7 and Staphylococcus aureus. Antagonistic activity was assayed by agar spot test and revealed that strain L. plantarum 105 had the strongest ability to contrast the growth of L. monocytogenes, while strains L. plantarum 106 and 107 were the most active microorganisms against E. coli O157:H7. The antimicrobial ability was also screened by well diffusion assay and broth micro-dilution method using cell-free supernatants (CFS) from each Lactobacillus strain. Moreover, the chemical nature of the molecules released in the CFS, and possibly underlying the antagonistic activity, was preliminary characterized by exposure to different constraints such as pH neutralization, heating, catalase, and proteinase treatments. Our data suggest that the ability of L. plantarum cultures to contrast pathogens growth in vitro depends, at least in part, on a pH-lowering effect of supernatants and/or on the presence of organic acids. Cluster analysis was performed in order to group L. plantarum strains according to their antimicrobial effect

  5. Methods for the Evaluation of the Bioactivity and Biocontrol Potential of Species of Trichoderma.

    Science.gov (United States)

    Steyaert, Johanna; Hicks, Emily; Kandula, Janaki; Kandula, Diwakar; Alizadeh, Hossein; Braithwaite, Mark; Yardley, Jessica; Mendoza-Mendoza, Artemio

    2016-01-01

    Members of the genus Trichoderma comprise the majority of commercial fungal biocontrol agents of plant diseases. As such, there is a wealth of information available on the analysis of their biocontrol potential and the mechanisms behind their superior abilities. This chapter aims to summarize the most common methods utilized within a Trichoderma biocontrol program for assessing the biological properties of individual strains. PMID:27565489

  6. Isolation and characterization of Bacillus subtilis EB-28, an endophytic bacterium strain displaying biocontrol activity against Botrytis cinerea Pers

    Institute of Scientific and Technical Information of China (English)

    Shutong WANG; Tongle HU; Yanling JIAO; Jianjian WEI; Keqiang CAO

    2009-01-01

    The fungal pathogen Botrytis cinerea Pers. causes severe rotting on tomato fruits during storage and shelf life. As a biological control agent, endophytic bacterium was regarded as an effective alternative to chemical control. Out of 238 endophytic bacterial isolates, three strains (EB-15, EB-28, and EB-122) isolated from Lycopersicum esculentum Mill., Speranskia tuberculata (Bge.) Baill, and Dictamnus dasycarpus Turcz. respectively were found to be strongly antagonistic to the pathogen in vitro and were selected for further in vivo tests. One endophytic bacterium strain, encoded EB-28, was selected from the three in vivo tested isolates. The inhibitive rate of EB-28 reached 71.1% in vitro and 52.4% in vivo. EB-28 was identified as Bacillus subtilis according to its morphological, physiological, and biochemical characteristics and 16S rDNA sequence analysis.

  7. Purification and characterization of an extracellular protease from Clonostachys rosea

    Institute of Scientific and Technical Information of China (English)

    LI Jun; HUANG Xiao-wei; ZHANG Ke-qin

    2004-01-01

    @@ An extracellular protease from Clonostachys rosea (syn. Gliocladium roseum) was purified to SDSPAGE homogeneity with 14-fold purification by ultrafiltration、 ammonium sulfate precipetation、hydrophobic interaction chromatography and anion exchange chromatography. The molecular weight of the protease was 32 kDa as estimated by SDS-PAGE. The N-terminal sequence of first 10 amino acids was A-T-Q-S-N-A-P-W-G-L. This enzyme exhibited pH and temperature optima of 9-10 and 60℃, respectively, and was stable over a wide range of pH 4-10 and temperature 4-50 ℃. It did not require Ca2+ for activity and thermal stability. Pre-incubation of enzyme with Zn2+ , Cu2+ , Hg2+,Fe3+ inhibited most of the enzyme activity, but Mn2+ increased enzyme activity up to 38%. It remained stable in the presence of Tween20, H2O2, EDTA. The inhibition profile of the enzymes by PMSF, suggested that this purified protease belongs to the serine protease family. The protease could immobilize nematodes (Panagrellus redivirus) in bioassays and hydrolyzed proteins of the purified cuticle.

  8. Biocontrol Effects of Ginseng Rust Rot by Two Strains of Streptomyces spp%2株放线菌对人参锈腐病的生物防治效果

    Institute of Scientific and Technical Information of China (English)

    周淑香; 张连学; 鲁新; 李小宇; 李玉

    2012-01-01

    [Objective] The aim was to explore biocontrol effects of ginseng rust rot by two strains of Streptomyces spp.. [ Method] The antagonistic action of two strains of Streptomyces spp. Selected from rhizosphere of ginseng on Cylindrocarpon destructans was studied by indoor antagonistic test. The control effect of two strains of Streptomyces spp. Against ginseng rust rot as well as effects of two strains of Streptomyces spp. On the yield and growth of ginseng were studied by field biocontrol test. [ Result] The two strains of Streptomyces spp. Both had stronger inhibition to Cylindrocarpon destructans. After the two strains of Streptomyces spp. Were applied, disease indexes of ginseng rust rot both significantly decreased (P < 0.01) ,and their control effects reached 41.94% and 44.28% ,respectively. Compared with control,the yield of ginseng treated by two strains of Streptomyces spp. Were higher,but the difference was not significant. The two strains of Streptomyces spp. Had no significant effect on the survival rate of ginseng seedings and plant growth. [Conclusion] The research provides theoretical basis for studies on the biocontrol of ginseng rust rot.%[目的]探讨放线菌对人参锈腐病的生物防治效果.[方法]通过室内拮抗试验,研究了从人参根际分离出来的2株放线菌对人参锈腐菌(Cylindrocarpon destructans)的拮抗作用;通过田间生防试验,测定了2株放线菌对人参锈腐病的防效及对人参产量和植株生长的影响.[结果]2株放线菌对人参锈腐菌均有较强的抑制作用.施用放线菌后人参锈腐病的病情指数显著降低(P<0.01),防治效果分别达41.94%和44.28%.施用放线菌后人参的产量与对照相比有所增加,但未达到显著水平;放线菌对参根成活率、株高和茎粗影响不显著.[结论]为人参锈腐病的生物防治研究提供了理论依据.

  9. 中国新记录属——枝穗霉属(丝孢纲)%The finding of the genus Clonostachys (Hyphomycetes) in China

    Institute of Scientific and Technical Information of China (English)

    高晓梅; 陈顺; 吕国忠; 李莉; 孙晓东; 刘晓辉; 朱万芹

    2008-01-01

    Clonostachys phyllophila and Clonostachys rogersoniana were isolated from rhizosphere soil of Allium tuberosum in Chaoyang, Liaoning Province. It is the first report of the genus in China. The species was described morphologically and illustrated in this paper, the culture was stored in the Culture Collection of Liaoning Center of Culture Collection (LCCC).

  10. Optimization of culture conditions of the biocontrol strain CNY-04 against Botryis cinerea by response surface methodology%响应面法优化灰霉病生防菌CNY-04培养条件

    Institute of Scientific and Technical Information of China (English)

    陆继臣; 迟乃玉; 窦少华; 奥杰

    2013-01-01

    [Objective] In order to raising the antagonistic effect,the growth conditions of biocontrol strain CNY-04 was optimized.[Methods] The growth conditions of the biocontrol strain CNY-04 were optimized by response surface methodology basing on single factor tests.The growth curve was also measured.[Results] The optimal culture conditions of the biocontrol strain CNY-04 were determined for 0.5% glucose,2.0% peptone,0.1% yeast extract,0.5% beef extract,temperature at 32 ℃,pH 8.0,speeding at 150 r/min,the incubation time of 48 h,inoculation amount of 4%,liquid volume of 75 mL/250 mL.The OD600 of CNY-04 was 2.907 with the optimal growth conditions,consistenting with the predicted value.The antibacterial circle diameter was 44.5 mm,improving by 30.9%.[Conclusion] The optimal growth condition of the biocontrol strain CNY-04 was identified,providing a theoretical basis for enlargement of production.%[目的]通过优化生防菌CNY-04的培养条件,提高其对灰霉病菌的抑菌效果.[方法]在单因素试验的基础上,利用响应面法(Response surface methodology)对灰霉病生防菌CNY-04培养条件进行整体优化,并测其生长曲线.[结果]生防菌CNY-04最优培养条件为牛肉膏0.5%、蛋白胨2.0%、酵母膏0.1%、葡萄糖0.5%、时间48 h、接种量4%、温度32℃、pH 8.0、装液量75 mL/250 mL和转速150 r/min,在此培养条件下生防菌CNY-04的OD600为2.907,与模型预测值相符,对灰葡萄孢菌的抑菌圈直径为44.5 mm,较优化前提高了30.9%.[结论]从整体上确定了生防菌CNY-04的最优培养条件,为该菌扩大化生产提供理论基础.

  11. Use of a non-homologous end-joining-deficient strain (delta-ku70) of the biocontrol fungus Trichoderma virens to investigate the function of the laccase gene lcc1 in sclerotia degradation.

    Science.gov (United States)

    Catalano, Valentina; Vergara, Mariarosaria; Hauzenberger, Jasmin R; Seiboth, Bernhard; Sarrocco, Sabrina; Vannacci, Giovanni; Kubicek, Christian P; Seidl-Seiboth, Verena

    2011-02-01

    The aim of this study was to apply a generated Δtku70 strain with increased homologous recombination efficiency from the mycoparasitic fungus Trichoderma virens for studying the involvement of laccases in the degradation of sclerotia of plant pathogenic fungi. Inactivation of the non-homologous end-joining pathway has become a successful tool in filamentous fungi to overcome poor targeting efficiencies for genetic engineering. Here, we applied this principle to the biocontrol fungus T. virens, strain I10, by deleting its tku70 gene. This strain was subsequently used to delete the laccase gene lcc1, which we found to be expressed after interaction of T. virens with sclerotia of the plant pathogenic fungi Botrytis cinerea and Sclerotinia sclerotiorum. Lcc1 was strongly upregulated at early colonization of B. cinerea sclerotia and steadily induced during colonization of S. sclerotiorum sclerotia. The Δtku70Δlcc1 mutant was altered in its ability to degrade the sclerotia of B. cinerea and S. sclerotiorum. Interestingly, while the decaying ability for B. cinerea sclerotia was significantly decreased, that to degrade S. sclerotiorum sclerotia was even enhanced, suggesting the operation of different mechanisms in the mycoparasitism of these two types of sclerotia by the laccase LCC1. PMID:20872221

  12. Identification and characterization of antagonistie strain biocontrol of cucumber Botrytis cinere and inhbition properties%黄瓜灰霉病拮抗菌的筛选鉴定及其抑菌特性研究

    Institute of Scientific and Technical Information of China (English)

    师宝忠; 仇艳肖; 马金亮; 周竞; 麻耀华

    2012-01-01

    从石家庄地区多个蔬菜大棚的根际土壤分离筛选到一株高效生防菌F1-2,对黄瓜灰霉病菌的抑菌圈直径高达33mm。根据菌株F1-2的形态特征、生理生化特性以及16SrDNA序列对其进行鉴定,初步确定为枯草芽孢杆菌(Bacillus subtilis)。采用生长速率法对拮抗菌的抑菌谱进行测定,其对所选的16种病原菌均有很好的抑制作用。实验还对拮抗菌F1-2的抑菌特性进行了研究,菌株连续传代至10代抑菌性能保持不变,菌株的发酵液对热和酸碱都比较稳定。该菌株的代谢产物具有广谱抑菌活性和较强的稳定性,具有开发成为灰霉病生防制剂的潜力。%An effective fungal strain F1--2 was isolated from rhizosphere soil of several greenhouses in Shiji- azhuang. The strain suppressed the growth of botrytis cinerea which was isolated from cucumber with the inhibition zone diameter of 33mm. The physiological and biochemical characteristics of the strain F1--2 and the identity of the 16SrDNA point out that this organism is Bacillus subtilis. Antimicrobial spectrum of the strain ,which could restrain the growth of 16 pathogens,was determined by growth rate method. The inhi- bition properties of strain F1--2 was researched in the experiment. After 10 generations,the antagonistic activity of the strain kept stable. Fermentation liquid of the strains have stability to heat, acid and base Metabolite of the strains was able to inhibit many kinds of pathogens and have high stability,which laid the foundation of development as botrytis cinerea suppressing Bio--control preparations.

  13. Identification of a novel small cysteine-rich protein in the fraction from the biocontrol Fusarium oxysporum strain CS-20 that mitigates Fusarium wilt symptoms and triggers defense responses in tomato

    Directory of Open Access Journals (Sweden)

    Larisa A. Shcherbakova

    2016-01-01

    Full Text Available The biocontrol effect of the nonpathogenic F. oxysporum strain CS-20 against the tomato wilt pathogen F. oxysporum f. sp. lycopersici (FOL has been previously reported to be primarily plant-mediated. This study shows that CS-20 produces proteins, which elicit defense responses in tomato plants. Three protein-containing fractions were isolated from CS-20 biomass using size exclusion chromatography. Exposure of seedling roots to one of these fractions prior to inoculation with pathogenic FOL strains significantly reduced wilt severity. This fraction initiated an ion exchange response in cultured tomato cells resulting in a reversible alteration of extracellular pH; increased tomato chitinase activity, and induced systemic resistance by enhancing PR-1 expression in tomato leaves. Two other protein fractions were inactive in seedling protection. The main polypeptide (designated CS20EP, which was specifically present in the defense-inducing fraction and was not detected in inactive protein fractions, was identified. The nucleotide sequence encoding this protein was determined, and its complete amino acid sequence was deduced from direct Edman degradation (25 N-terminal amino acid residues and DNA sequencing. The CS20EP was found to be a small basic cysteine-rich protein with a pI of 9.87 and 23.43% of hydrophobic amino acid residues. BLAST search in the NCBI database showed that the protein is new; however, it displays 48% sequence similarity with a hypothetical protein FGSG_10784 from F. graminearum strain PH-1. The contribution of CS20EP to elicitation of tomato defense responses resulting in wilt mitigating is discussed.

  14. Characterization of a novel plant growth-promoting bacteria strain Delftia tsuruhatensis HR4 both as a diazotroph and a potential biocontrol agent against various plant pathogens.

    Science.gov (United States)

    Han, Jigang; Sun, Lei; Dong, Xiuzhu; Cai, Zhengqiu; Sun, Xiaolu; Yang, Hailian; Wang, Yunshan; Song, Wei

    2005-01-01

    A novel, plant growth-promoting bacterium Delftia tsuruhatensis, strain HR4, was isolated from the rhizoplane of rice (Oryza sativa L., cv. Yueguang) in North China. In vitro antagonistic assay showed this strain could suppress the growth of various plant pathogens effectively, especially the three main rice pathogens (Xanthomonas oryzae pv. oryzae, Rhizoctonia solani and Pyricularia oryzae Cavara). Treated with strain HR4 culture, rice blast, rice bacterial blight and rice sheath blight for cv. Yuefu and cv. Nonghu 6 were evidently controlled in the greenhouse. Strain HR4 also showed a high nitrogen-fixing activity in N-free Döbereiner culture medium. The acetylene reduction activity and 15N2-fixing activity (N2FA) were 13.06 C2H4 nmolml(-1) h(-1) and 2.052 15Na.e.%, respectively. The nif gene was located in the chromosome of this strain. Based on phenotypic, physiological, biochemical and phylogenetic studies, strain HR4 could be classified as a member of D. tsuruhatensis. However, comparisons of characteristics with other known species of the genus Delftia suggested that strain HR4 was a novel dizotrophic PGPB strain. PMID:15709367

  15. Exploitation of gene(s) involved in 2,4-diacetylphloroglucinol biosynthesis to confer a new biocontrol capability to a Pseudomonas strain.

    OpenAIRE

    Fenton, A M; Stephens, P M; Crowley, J; O'Callaghan, M; O'Gara, F.

    1992-01-01

    Tn5 mutagenesis and complementation analysis were used to clone a 6-kb genomic fragment required for biosynthesis of 2,4-diacetylphloroglucinol (Phl) from fluorescent Pseudomonas sp. strain F113. A recombinant plasmid, pCU203, containing this region partially complemented a Phl production-negative mutant (F113G22) derived from strain F113. When sugar beet seeds were sown into an unsterilized soil, in which sugar beet was subject to damping-off by Pythium ultimum, the emergence of sugar beet s...

  16. Biocontrol of Rhizoctonia solani with Trichoderma Spp.

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@ From over 800 fungal strains of Trichoderma Spp. , 6 strains were found to greatly inhibit the growing of Rhizocotonia solani, the pathogen of rice sheath blight in dual culture. Among them, strain T3 was the best antagonist,which reduced the growing of the pathogen by 52.54% (Table 1). In field, both the pesticide Jinggangmycin and the mixture of T1 T6 could reduce the severity of rice sheath blight(Table 2), which resulted in the increases of seed setting rate and 1000 grain weight. Because the effect of the antagonists on the control of the pathogen could be partially realized in the watery environment, studies on the biocontrol mechanism of the fungi should be strengthened to help the establishment of a best way of antagonist utilization.

  17. Some fungicides and growth inhibitor/biocontrol-enhancer 2-deoxy-D-glucose increase patulin from penicillium expansum strains in vitro

    OpenAIRE

    Paterson, R. R. M.

    2007-01-01

    This study assessed the effect of disease control agents, fungicides and 2-deoxy-D-glucose (DOG), on a mycotoxin, patulin, production by strains of Penicillium expansum. Fungicides and DOG were added individually to agar on which the fungus was then grown. In an initial screen of six fungicides, three indicated that patulin was stimulated. These were tested further at a range of concentrations and colony diameters (cd) and patulin production were measured. Although patulin production appe...

  18. Expression of a serine protease gene prC is up-regulated by oxidative stress in the fungus Clonostachys rosea: implications for fungal survival.

    Directory of Open Access Journals (Sweden)

    Cheng-Gang Zou

    Full Text Available BACKGROUND: Soil fungi face a variety of environmental stresses such as UV light, high temperature, and heavy metals. Adaptation of gene expression through transcriptional regulation is a key mechanism in fungal response to environmental stress. In Saccharomyces cerevisiae, the transcription factors Msn2/4 induce stress-mediated gene expression by binding to the stress response element. Previous studies have demonstrated that the expression of extracellular proteases is up-regulated in response to heat shock in fungi. However, the physiological significance of regulation of these extracellular proteases by heat shock remains unclear. The nematophagous fungus Clonostachys rosea can secret an extracellular serine protease PrC during the infection of nematodes. Since the promoter of prC has three copies of the stress response element, we investigated the effect of environmental stress on the expression of prC. METHODOLOGY/PRINCIPAL FINDINGS: Our results demonstrated that the expression of prC was up-regulated by oxidants (H(2O(2 or menadione and heat shock, most likely through the stress response element. After oxidant treatment or heat shock, the germination of conidia in the wild type strain was significantly higher than that in the prC mutant strain in the presence of nematode cuticle. Interestingly, the addition of nematode cuticle significantly attenuated the production of reactive oxygen species (ROS induced by oxidants and heat shock in the wild type strain, but not in prC mutant strain. Moreover, low molecule weight (<3 kD degradation products of nematode cuticle suppressed the inhibitory effect of conidial germination induced by oxidants and heat shock. CONCLUSIONS/SIGNIFICANCE: These results indicate that PrC plays a protective role in oxidative stress in C. rosea. PrC degrades the nematode cuticle to produce degradation products, which in turn offer a protective effect against oxidative stress by scavenging ROS. Our study reveals a novel

  19. Novel routes for improving biocontrol activity of Bacillus based bioinoculants

    Directory of Open Access Journals (Sweden)

    Liming eWu

    2015-12-01

    Full Text Available Biocontrol formulations prepared from plant-growth-promoting bacteria are increasingly applied in sustainable agriculture. Especially inoculants prepared from endospore-forming Bacillus strains have been proven as efficient and environmental-friendly alternative to chemical pesticides due to their long shelf life, which is comparable with that of agrochemicals. However, these formulations of the first generation are sometimes hampered in their action and do not fulfill in each case the expectations of the appliers. In this review we use the well-known plant-associated Bacillus amyloliquefaciens type strain FZB42 as example for the successful application of different techniques offered today by comparative, evolutionary and functional genomics, site-directed mutagenesis and strain construction including marker removal, for paving the way for preparing a novel generation of biocontrol agents.

  20. Attachment capability of antagonistic yeast Rhodotorula glutinis to Botrytis cinerea contributes to biocontrol efficacy

    Directory of Open Access Journals (Sweden)

    Boqiang eLi

    2016-05-01

    Full Text Available Rhodotorula glutinis as an antagonism show good biocontrol performance against various postharvest diseases in fruits. In the present study, strong attachment capability of R. glutinis to spores and hyphae of Botrytis cinerea was observed. Further analysis showed that certain protein components on the yeast cell surface played critical role during the interaction between R. glutinis and B. cinerea. The components mainly distributed at the poles of yeast cells and might contain glycosylation modification, as tunicamycin treated yeast cells lost attachment capability to B. cinerea. To investigate contributions of attachment capability of R. glutinis to its biocontrol efficacy, yeast cells were mutagenized with 3% methane-sulfonic acid ethyl ester (EMS, and a mutant CE4 with stable non-attaching phenotype was obtained. No significant difference was found on colony, cell morphology, reproductive ability and capsule formation between the mutant and wild type. However, there was a distinct difference in India ink positive staining patterns between the two strains. Moreover, wild type strain of R. glutinis showed better performance on inhibiting spore germination and mycelial growth of B. cinerea than CE4 strain when yeast cells and B. cinerea were co-cultured in vitro. In biocontrol assay, both wild type and CE4 strains showed significant biocontrol efficacy against gray mould caused by B. cinerea in apple fruit, whereas, control effect of CE4 strain was lower than that of wild type. Our findings provided new evidences that attachment capability of R. glutinis to B. cinerea contributed to its biocontrol efficacy.

  1. Identification of mycoparasitism-related genes in Clonostachys rosea 67-1 active against Sclerotinia sclerotiorum.

    Science.gov (United States)

    Sun, Zhan-Bin; Sun, Man-Hong; Li, Shi-Dong

    2015-01-01

    Clonostachys rosea is a mycoparasite that has shown great potential in controlling various plant fungal pathogens. In order to find mycoparasitism-related genes in C. rosea, the transcriptome of the efficient isolate 67-1 in association with sclerotia of Sclerotinia sclerotiorum was sequenced and analysed. The results identified 26,351 unigenes with a mean length of 1,102 nucleotides, among which 18,525 were annotated in one or more databases of NR, KEGG, Swiss-Prot, GO and COG. Differentially expressed genes at 8 h, 24 h and 48 h after sclerotial induction were analysed, and 6,890 unigenes were upregulated compared with the control without sclerotia. 713, 1,008 and 1,929 genes were specifically upregulated expressed, while 1,646, 283 and 529 genes were specifically downregulated, respectively. Gene ontology terms analysis indicated that these genes were mainly involved in metabolism of biological process, catalysis of molecular function and cellular component. The expression levels of 12 genes that were upregulated after encountering with S. sclerotiorum were monitored using real-time PCR. The results indicated that the quantitative detection was consistent with the transcriptome analysis. The study provides transcriptional gene expression information on C. rosea parasitizing S. sclerotiorum and forms the basis for further investigation of mycoparasitism-related genes of C. rosea. PMID:26657839

  2. Impact of 2,4-Diacetylphloroglucinol-Producing Biocontrol Strain Pseudomonas fluorescens F113 on Intraspecific Diversity of Resident Culturable Fluorescent Pseudomonads Associated with the Roots of Field-Grown Sugar Beet Seedlings

    OpenAIRE

    Moënne-Loccoz, Yvan; Tichy, Hans-Volker; O'Donnell, Anne; Simon, Reinhard; O'Gara, Fergal

    2001-01-01

    The impact of the 2,4-diacetylphloroglucinol-producing biocontrol agent Pseudomonas fluorescens F113Rif on the diversity of the resident community of culturable fluorescent pseudomonads associated with the roots of field-grown sugar beet seedlings was evaluated. At 19 days after sowing, the seed inoculant F113Rif had replaced some of the resident culturable fluorescent pseudomonads at the rhizoplane but had no effect on the number of these bacteria in the rhizosphere. A total of 498 isolates ...

  3. Surfactin triggers biofilm formation of Bacillus subtilis in melon phylloplane and contributes to the biocontrol activity.

    Science.gov (United States)

    Zeriouh, Houda; de Vicente, Antonio; Pérez-García, Alejandro; Romero, Diego

    2014-07-01

    The biocontrol activity of many Bacillus species has been traditionally related to the direct antagonism of pathogens. In previous works, we reported that B. subtilis strain UMAF6614 was an efficient biocontrol agent that produced bacillomycin, fengycin and surfactin lipopeptides. Bacillomycins and fengycins were shown to have antagonistic activity towards fungal and bacterial pathogens of cucurbits; however, the functionality of surfactin remained unclear. In this study, the role of surfactin in the biocontrol activity of this strain was investigated. We observed that a deficiency in surfactin production led to a partial reduction of disease suppression by this biocontrol agent, which coincided with a defect in biofilm formation and the colonization of the melon phylloplane. These effects were due to a dramatic reduction in the production of exopolysaccharide and the TasA protein, which are the two major components of the extracellular matrix. We propose that the biocontrol activity of this strain is the result of the coordinated action of the three families of lipopeptides. B. subtilis UMAF6614 produces surfactin to trigger biofilm formation on melon phylloplane, which ensures the long-term persistence and the adequate secretion of suppressive lipopeptides, bacillomycins and fengycins, which efficiently target pathogens.

  4. Whole genome shotgun sequence of Bacillus amyloliquefaciens TF28, a biocontrol entophytic bacterium.

    Science.gov (United States)

    Zhang, Shumei; Jiang, Wei; Li, Jing; Meng, Liqiang; Cao, Xu; Hu, Jihua; Liu, Yushuai; Chen, Jingyu; Sha, Changqing

    2016-01-01

    Bacillus amyloliquefaciens TF28 is a biocontrol endophytic bacterium that is capable of inhibition of a broad range of plant pathogenic fungi. The strain has the potential to be developed into a biocontrol agent for use in agriculture. Here we report the whole-genome shotgun sequence of the strain. The genome size of B. amyloliquefaciens TF28 is 3,987,635 bp which consists of 3754 protein-coding genes, 65 tandem repeat sequences, 47 minisatellite DNA, 2 microsatellite DNA, 63 tRNA, 7rRNA, 6 sRNA, 3 prophage and CRISPR domains. PMID:27688836

  5. Bacteriophage biocontrol of foodborne pathogens.

    Science.gov (United States)

    Kazi, Mustafa; Annapure, Uday S

    2016-03-01

    Bacteriophages are viruses that only infect bacterial cells. Phages are categorized based on the type of their life cycle, the lytic cycle cause lysis of the bacterium with the release of multiple phage particles where as in lysogenic phase the phage DNA is incorporated into the bacterial genome. Lysogeny does not result in lysis of the host. Lytic phages have several potential applications in the food industry as biocontrol agents, biopreservatives and as tools for detecting pathogens. They have also been proposed as alternatives to antibiotics in animal health. Two unique features of phage relevant for food safety are that they are harmless to mammalian cells and high host specificity, keeping the natural microbiota undisturbed. However, the recent approval of bacteriophages as food additives has opened the discussion about 'edible viruses'. This article reviews in detail the application of phages for the control of foodborne pathogens in a process known as "biocontrol". PMID:27570260

  6. Screening of High-performance Biocontrol Strains of Nematode-trapping Fungi from Yak Dung in Plateau Pastures of Yunnan Province%云南高海拔牧场牦牛捕食线虫真菌生防高效菌株的筛选

    Institute of Scientific and Technical Information of China (English)

    舒凡帆; 代琦; 苏锡钧; 苏子峰; 苏鸿雁

    2014-01-01

    目的:为筛选云南高海拔牧场牦牛高效力的寄生线虫生防真菌菌株。方法:对云南大理花甸坝牧场,中甸打日坝牧场及白马雪山牧场采集的少孢节丛孢(A. oligospora)、弯孢节丛孢(A. musiformis)、椭圆隔指孢(D. ellipsospora)和长孢隔指孢(D. leptospora)采用盐酸胃蛋白酶处理筛选耐受菌株,并进一步比较了耐受菌株的捕食活力。结果:12株捕食线虫真菌中,有6株通过盐酸胃蛋白酶处理,通过率为50%,其中大理花甸坝和中甸打日坝的A. oligospora及白马雪山牧场的D. ellipsospora显示出较强的捕食活力。结论:本研究初步筛选出3株生防高效菌株,为牦牛及其它动物线虫病的生物防治积累了捕食线虫真菌菌种资源。%Objective: To screen high efficiency biocontrol strains of nematode-trapping fungi isolated from yak dung in plateau pastures of Yunnan Province. Methods:Twelve nematode-trapping fungi of A. oligospora, A. musiformis, D. ellipsospora, D. leptospora isolated from Dali Huadianba pasture, Zhongdian Dariba pasture, Baima Snow Mountain pasture were tested in pepsin-hydrochloric acid and the nematode-trapping activity was determined subsequently. Results:Half of isolates survived in pepsin-hydrochloric acid treatment and three isolates A. oligospora from Huadianba pasture and Dariba pasture and D. ellipsospora from Baima Snow Mountain pasture displayed high nematode-trapping activity under laboratory conditions. Conclusion: Three high efficiency biocontrol nematode-trapping fungi were obtained and this research accumulated biocontrol strain resources in the biological control of nematode disease of yak and other animals.

  7. Postharvest Biocontrol: Introspection and Paradigm Shifts

    Science.gov (United States)

    The use of postharvest biocontrol agents as an alternative to the synthetic, chemical fungicides on a widespread basis has many constraints. During the last twenty years, the field of postharvest biocontrol research has significantly grown and developed and seen the creation of several products. Des...

  8. Heat shock improves stress tolerance and biocontrol performance of Rhodotorula mucilaginosa

    Science.gov (United States)

    Biological control of postharvest diseases, utilizing wild species and strains of antagonistic yeasts, is a research topic and eco-friendly management approach that has received considerable attention. A few yeast-based biocontrol products are available in the market. Ecological fitness, stress to...

  9. Pantoea applied genomics to understand and improve biocontrol activity against fire blight

    Science.gov (United States)

    Pantoea agglomerans and P. vagans (ex. Erwinia herbicola) are common epiphytes of pome fruit flowers and three strains (E325, P10c, C9-1) have been commercially developed as effective biocontrol products for managing fire blight (Erwinia amylovora). Antibiotics as a standard, reliable chemical optio...

  10. Pichia angusta is an effective biocontrol yeast against postharvest decay of apple fruit caused by Botrytis cinerea and Monilia fructicola.

    Science.gov (United States)

    Fiori, Stefano; Fadda, Angela; Giobbe, Sara; Berardi, Enrico; Migheli, Quirico

    2008-09-01

    The efficacy of eight isolates of Pichia angusta against three common postharvest pathogens of apple fruit was evaluated for the first time. All tested strains showed significant biocontrol activity against both Botrytis cinerea and Monilia fructicola, whereas efficacy against Penicillium expansum was poor. A leucine-auxotrophic mutant had no significant biocontrol activity against brown rot of apple, while the addition of 0.6-1.2 g L(-1) leucine in the fruit wound fully restored the biocontrol activity of this mutant against M. fructicola. Given the extremely well-developed classical and molecular genetics, the availability of genomic libraries, and its complete genomic sequence, this species can serve to elucidate the mechanisms related to biocontrol capacity.

  11. Attachment Capability of Antagonistic Yeast Rhodotorula glutinis to Botrytis cinerea Contributes to Biocontrol Efficacy.

    Science.gov (United States)

    Li, Boqiang; Peng, Huaimin; Tian, Shiping

    2016-01-01

    Rhodotorula glutinis as an antagonism show good biocontrol performance against various post-harvest diseases in fruits. In the present study, strong attachment capability of R. glutinis to spores and hyphae of Botrytis cinerea was observed. Further analysis showed that certain protein components on the yeast cell surface played critical role during the interaction between R. glutinis and B. cinerea. The components mainly distributed at the poles of yeast cells and might contain glycosylation modification, as tunicamycin treated yeast cells lost attachment capability to B. cinerea. To investigate contributions of attachment capability of R. glutinis to its biocontrol efficacy, yeast cells were mutagenized with 3% methane-sulfonic acid ethyl ester (EMS), and a mutant CE4 with stable non-attaching phenotype was obtained. No significant difference was found on colony, cell morphology, reproductive ability, and capsule formation between the mutant and wild-type. However, there was a distinct difference in India ink positive staining patterns between the two strains. Moreover, wild-type strain of R. glutinis showed better performance on inhibiting spore germination and mycelial growth of B. cinerea than CE4 strain when yeast cells and B. cinerea were co-cultured in vitro. In biocontrol assay, both wild-type and CE4 strains showed significant biocontrol efficacy against gray mold caused by B. cinerea in apple fruit, whereas, control effect of CE4 strain was lower than that of wild-type. Our findings provided new evidences that attachment capability of R. glutinis to B. cinerea contributed to its biocontrol efficacy. PMID:27199931

  12. Impact of a Recombinant Biocontrol Bacterium, Pseudomonas fluorescens pc78, on Microbial Community in Tomato Rhizosphere

    Directory of Open Access Journals (Sweden)

    Hyun Gi Kong

    2016-04-01

    Full Text Available Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer.

  13. Impact of a Recombinant Biocontrol Bacterium, Pseudomonas fluorescens pc78, on Microbial Community in Tomato Rhizosphere.

    Science.gov (United States)

    Kong, Hyun Gi; Kim, Nam Hee; Lee, Seung Yeup; Lee, Seon-Woo

    2016-04-01

    Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer. PMID:27147933

  14. 小麦孢囊线虫病生防真菌08F04菌株的鉴定及防效测定%Identification and Effects of Biocontrol Fungal Strain 08F04 on Cereal Cyst Nematode of Wheat

    Institute of Scientific and Technical Information of China (English)

    张洁; 袁虹霞; 孙炳剑; 陈莉; 付博; 李洪连

    2013-01-01

    小麦禾谷孢囊线虫病已成为我国黄淮麦区的重要病害,目前生产上尚缺乏有效的防控方法。菌株08F04是作者从禾谷孢囊线虫孢囊上分离的1株寄生真菌。本研究通过形态学和分子生物学方法,将其鉴定为球孢白僵菌Beauveria bassiana (Bals.) Vuill。2011和2012年对该菌株进行了盆栽和田间防效试验,菌株08F04麦粒砂培养物6%(w/w)土壤处理的盆栽试验孢囊减退率分别达到65.03%和50.95%;田间试验中,菌剂08F0420 g·m−2沟施处理土壤,2011年在许昌和荥阳的孢囊减退率分别为58.28%和32.13%,2012年分别达到58.49%和44.50%。结果表明,菌株08F04是防治小麦禾谷孢囊线虫病具有较大潜在开发价值的生防菌株。%Cereal cyst nematode (CCN) has become a serious disease in Huanghuai wheat production area. The biocontrol strain 08F04 is parasitic fungus which was isolated from the cyst in the diseased fields. It was identified as Beauveria bassiana (Bals.) Vuill by morphology and rDNA-ITS sequence analyses. Its control efficiency on CCN was evaluated in the greenhouse and diseased fields (Xuchang and Xingyang), respectively. The cysts decreasing rates were 65.03%and 50.95%respectively in 2011 and 2012, when the strain 08F04 mixed with soil (6% w/w) in pots. In field trial, the cysts decreasing rates in 2011 were 58.28% and 32.13% in Xingyang and Xuchang respectively, and in 2012 were 58.49% and 44.50% respectively. The fungal strain 08F04 could be supposed as a potential biocontrol agent of cereal cyst nematode of wheat.

  15. nematicidal activities and identification of a biocontrol bacterium YC-10 strain%生防细菌YC-10的杀线虫活性及其鉴定

    Institute of Scientific and Technical Information of China (English)

    成飞雪; 张德咏; 刘勇; 王忠勇; 程菊娥; 何明远; 肖启明

    2011-01-01

    A biocontrol bacterium YC-10 was isolated from tobacco and tested for the nematicidal activity. The bacterium culture diluted 1, 5, 10, 20 and 40 times all showed high nematicidal activity to Meloidogyne incognita and Heterodera glycines Ichinohe. After the nematodes were treated with the culture for 96 hours, the lethal rate of the M. incognita attained 100% and the H. glycines Ichinohe over 90%. The results indicated the potential application of biocontrol to nematode. The bacterium YC-10 was identified as Bacillus thuringiensis according to its 16S rDNA sequence and morphological, cultural, physiological and biochemical features. Study on the active substances of physical and chemical properties could showed that the substances were heat-stable and high temperature or the Proteinase K digestion not affect its nematicidal activity. The study also showed that nematicidal activity was stronger in acidic solution than in alkaline solution. The study is the foundation for separating the active ingredient and its mechanism of nematicidal activity.%从烟草中分离出 1株对根结线虫和胞囊线虫有较高生物活性的内生细菌菌株-YC-10.将该细菌发酵滤液原液及5、10、20和40倍稀释液处理南方根结线虫,96h后线虫死亡率均达到100%,处理大豆胞囊线虫96h,致死率大于90%,在植物线虫生物防治上显示出巨大的应用前景.通过形态特点、理化特征以及16SrDNA序列同源性分析将菌株YC-10鉴定为苏云金芽孢杆菌(Bacillus thuringiensis).对发酵液活性成分理化性质研究表明,活性成分对热稳定,高温、蛋白酶K处理不影响其杀虫活性,而酸性条件下杀线虫活性比碱性中更强.为后续杀线虫活性成分的分离及杀线虫机理研究奠定基础.

  16. Cucumber Rhizosphere Microbial Community Response to Biocontrol Agent Bacillus subtilis B068150

    OpenAIRE

    Lihua Li; Jincai Ma; A. Mark Ibekwe; Qi Wang; Ching-Hong Yang

    2015-01-01

    Gram-positive bacteria Bacillus subtilis B068150 has been used as a biocontrol agent against the pathogen Fusarium oxysporum cucumerinum. Cucumber was grown in three soils with strain B068150 inoculated in a greenhouse for 90 days, and the colonization ability of strain B068150 in cucumber rhizosphere and non-rhizosphere soils was determined. Changes in total bacteria and fungi community composition and structures using denaturing gradient gel electrophoresis (DGGE) and sequencing were determ...

  17. Transformation of Cyclaneusma minus with Green Fluorescent Protein (GFP to Enable Screening of Fungi for Biocontrol Activity

    Directory of Open Access Journals (Sweden)

    Alison Stewart

    2012-02-01

    Full Text Available Cyclaneusma needle-cast has a major impact on the New Zealand forest industry. The causal agent, Cyclaneusma minus, causes most severe damage to 11–20 year-old trees and currently there are no economically viable procedures for control of the disease in New Zealand. Here we present a method for genetic transformation of C. minus using protoplasts generated by incubation with Glucanex™ enzyme. C. minus was transformed with a gene encoding green fluorescent protein (GFP and expression was stable after successive sub-culturing of the strain in the absence of selective pressure. Expression of the gfp gene allowed us to utilize an in vitro GFP-based screening method to identify strains of Trichoderma with potential for biocontrol of this disease. The strain that showed the most promise as a potential biocontrol candidate exhibited a low level of inhibition by uncharacterized metabolite(s that C. minus secretes into the medium, and consistently caused a loss of GFP expression from the GFP-labeled C. minus strain. The interaction between C. minus and the biocontrol strain, in the interaction zone where GFP expression was lost, was determined to be fungicidal. The utility of such biocontrol strains is discussed. This study represents the first genetic manipulation of C. minus and will pave the way for further studies of the life cycle and infection biology of this organism.

  18. Draft Genome Sequences of Pseudomonas fluorescens Strains PA4C2 and PA3G8 and Pseudomonas putida PA14H7, Three Biocontrol Bacteria against Dickeya Phytopathogens.

    Science.gov (United States)

    Cigna, Jérémy; Raoul des Essarts, Yannick; Mondy, Samuel; Hélias, Valérie; Beury-Cirou, Amélie; Faure, Denis

    2015-01-29

    Pseudomonas fluorescens strains PA4C2 and PA3G8 and Pseudomonas putida strain PA14H7 were isolated from potato rhizosphere and show an ability to inhibit the growth of Dickeya phytopathogens. Here, we report their draft genome sequences, which provide a basis for understanding the molecular mechanisms involved in antibiosis against Dickeya.

  19. Draft Genome Sequences of Pseudomonas fluorescens Strains PA4C2 and PA3G8 and Pseudomonas putida PA14H7, Three Biocontrol Bacteria against Dickeya Phytopathogens

    OpenAIRE

    Cigna, Jérémy; Raoul des Essarts, Yannick; Mondy, Samuel; Hélias, Valérie; Beury-Cirou, Amélie; Faure, Denis

    2015-01-01

    Pseudomonas fluorescens strains PA4C2 and PA3G8 and Pseudomonas putida strain PA14H7 were isolated from potato rhizosphere and show an ability to inhibit the growth of Dickeya phytopathogens. Here, we report their draft genome sequences, which provide a basis for understanding the molecular mechanisms involved in antibiosis against Dickeya.

  20. Preliminary in vitro insights into the use of natural fungal pathogens of leaf-cutting ants as biocontrol agents.

    Science.gov (United States)

    Folgarait, Patricia; Gorosito, Norma; Poulsen, Michael; Currie, Cameron R

    2011-09-01

    Leaf-cutting ants are one of the main herbivores of the Neotropics, where they represent an important agricultural pest. These ants are particularly difficult to control because of the complex network of microbial symbionts. Leaf-cutting ants have traditionally been controlled through pesticide application, but there is a need for alternative, more environmentally friendly, control methods such as biological control. Potential promising biocontrol candidates include the microfungi Escovopsis spp. (anamorphic Hypocreales), which are specialized pathogens of the fungi the ants cultivate for food. These pathogens are suppressed through ant behaviors and ant-associated antibiotic-producing Actinobacteria. In order to be an effective biocontrol agent, Escovopsis has to overcome these defenses. Here, we evaluate, using microbial in vitro assays, whether defenses in the ant-cultivated fungus strain (Leucoagaricus sp.) and Actinobacteria from the ant pest Acromyrmex lundii have the potential to limit the use of Escovopsis in biocontrol. We also explore, for the first time, possible synergistic biocontrol between Escovopsis and the entomopathogenic fungus Lecanicillium lecanii. All strains of Escovopsis proved to overgrow A. lundii cultivar in less than 7 days, with the Escovopsis strain isolated from a different leaf-cutting ant species being the most efficient. Escovopsis challenged with a Streptomyces strain isolated from A. lundii did not exhibit significant growth inhibition. Both results are encouraging for the use of Escovopsis as a biocontrol agent. Although we found that L. lecanii can suppress the growth of the cultivar, it also had a negative impact on Escovopsis, making the success of simultaneous use of these two fungi for biocontrol of A. lundii questionable. PMID:21739253

  1. Non-aflatoxigenic Aspergillus flavus as potential biocontrol agents to reduce aflatoxin contamination in peanuts harvested in Northern Argentina.

    Science.gov (United States)

    Alaniz Zanon, María Silvina; Barros, Germán Gustavo; Chulze, Sofía Noemí

    2016-08-16

    Biological control is one of the most promising strategies for preventing aflatoxin contamination in peanuts at field stage. A population of 46 native Aspergillus flavus nonaflatoxin producers were analysed based on phenotypic, physiological and genetic characteristics. Thirty-three isolates were characterized as L strain morphotype, 3 isolates as S strain morphotype, and 10 isolates did not produce sclerotia. Only 11 of 46 non-aflatoxigenic isolates did not produce cyclopiazonic acid. The vegetative compatibility group (VCG) diversity index for the population was 0.37. For field trials we selected the non-aflatoxigenic A. flavus AR27, AR100G and AFCHG2 strains. The efficacy of single and mixed inocula as potential biocontrol agents in Northern Argentina was evaluated through a 2-year study (2014-2015). During the 2014 peanut growing season, most of the treatments reduced the incidence of aflatoxigenic strains in both soil and peanut kernel samples, and no aflatoxin was detected in kernels. During the 2015 growing season, there was a reduction of aflatoxigenic strains in kernel samples from the plots treated with the potential biocontrol agents. Reductions of aflatoxin contamination between 78.36% and 89.55% were observed in treated plots in comparison with the un-inoculated control plots. This study provides the first data on aflatoxin biocontrol based on competitive exclusion in the peanut growing region of Northern Argentina, and proposes bioproducts with potential use as biocontrol agents. PMID:27220011

  2. Recent advance of nematophagous fungi from China and with a special case on Clonostachys rosea (Gliocladium roseum)

    Institute of Scientific and Technical Information of China (English)

    ZHANG Ke-qin; ZHAO Ming-lan

    2004-01-01

    @@ China is one of the twelve prolific biological diversity countries in the world, and nematophagous fungol resources are very abundant. In this paper, the research advance of nematophagous fungi is discussed from six aspects: resources introduction, biology of nematophagous fungi, the application of biocontrol agents, fungistsis in soils, research on nematicidal metabolites and molecular biology on nematophagous fungi.

  3. Construction of new GFP-tagged fusants for Trichoderma harzianum with enhanced biocontrol activity

    Directory of Open Access Journals (Sweden)

    Kowsari Mojegan

    2014-07-01

    Full Text Available Trichoderma is one of the most exploited biocontrol agents for the management of plant diseases. In biocontrol ecology, the critical factors are detection, and the monitoring and recovery of specific biocontrol agents either naturally present or deliberately released into the environment. Protoplast fusion is an appropriate tool for the improvement of biocontrol Trichoderma strains. Protoplast isolation from Trichoderma harzianum was achieved using 24 h culture age, 6.6 mg/ml Novazym L 1412 at 30°C which resulted the maximum protoplast yield of 5 × 108/ml. The self-fused protoplasts were regenerated and 12 fusants were selected based on their growth rate on 2% colloidal chitin medium. Next, a comparison was done for chitinase and antagonistic activity. Transcriptomic analysis based on quantitative real-time RT-PCR, demonstrated that T8-05 fusant expressed 1.5 fold of chit42 transcript as compared with the parental line. This fusant with 7.02±0.15U chitinase activity showed a higher growth inhibition rate (100% than the parent strain, against Rhizoctonia solani. To obtain a genetically marked fusant that can be used as a biomonitor, the fusant was cotransformed with the gfp and amdS genes. The morphology and viability of selected cotransformant (FT8-7MK-05-2 was similar to the parent. Green fluorescing conidia were observed within the first 2 days of incubation in the soil, and this was followed by the formation of chlamydopores after 60 days. The colonisation of the gfp-tagged fusant was also monitored visually on R. solani sclerotia by scanning electron microscopy. Production of gfp-tagged fusant of Trichoderma spp. provides a potentially useful tool for monitoring hyphal growth patterns and the population of biocontrol agent isolates introduced into environmental systems.

  4. Isolation and Identification of an Antifungal Biocontrol Bacterium Strain Xj11%1株拮抗真菌的生防细菌Xj11的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    敖武; 曾庆桂; 李希茜; 李平; 邓映明; 朱笃

    2013-01-01

    An antagonistic activity bacterium strain Xj11 which had significantly antagonistic effect on plant pathogenic fungi,was selected from the disease Phyllostachys heterocycla CV.in Jinggangshan natural reserve,Jiangxi Province.The results of plate antagonistic experiment showed that strain Xj11 had strong antagonistic activities against Gibberella zeae,Fuarium oxysporum,Arthrinium sacchari,Alternaria alternate and Xylaria sp.According to the characteristics of morphology,physiology and biochemistry tests and 16S rDNA sequence analysis,the strain Xj11 was preliminary identified as Burkholderia gladioli strain Xj11.%从江西省井冈山自然保护区发病毛竹中分离到1株具有明显抑制植物病原真菌活性的生防细菌Xj11.其对小麦赤霉病菌、尖孢镰刀菌、甘蔗节菱孢菌、链格孢菌和炭角菌均有较强的抑制作用.通过形态特征和培养特征观察、生理生化实验及16S rDNA序列系统发育分析,初步鉴定该菌为唐菖蒲伯克霍尔德氏菌,命名为Burkholderia gladioli strain Xj11.

  5. Phytohormone profiles induced by Trichoderma isolates correspond with their biocontrol and plant growth-promoting activity on melon plants

    NARCIS (Netherlands)

    Martínez-Medina, Ainhoa; Del Mar Alguacil, Maria; Pascual, Jose A.; van Wees, Saskia C M

    2014-01-01

    The application of Trichoderma strains with biocontrol and plant growth-promoting capacities to plant substrates can help reduce the input of chemical pesticides and fertilizers in agriculture. Some Trichoderma isolates can directly affect plant pathogens, but they also are known to influence the ph

  6. Lactic acid bacteria from fresh fruit and vegetables as biocontrol agents of phytopathogenic bacteria and fungi

    OpenAIRE

    Trias Mansilla, Rosalia; Bañeras Vives, Lluís; Montesinos Seguí, Emilio; Badosa Romañó, Esther

    2008-01-01

    This study evaluated the efficacy of lactic acid bacteria (LAB) isolated from fresh fruits and vegetables as biocontrol agents against the phytopathogenic and spoilage bacteria and fungi, Xanthomonas campestris, Erwinia carotovora, Penicillium expansum, Monilinia laxa, and Botrytis cinerea. The antagonistic activity of 496 LAB strains was tested in vitro and all tested microorganisms except P. expansum were inhibited by at least one isolate. The 496 isolates were also analyzed for the inhibit...

  7. Biocontrol agents promote growth of potato pathogens, depending on environmental conditions.

    Science.gov (United States)

    Cray, Jonathan A; Connor, Mairéad C; Stevenson, Andrew; Houghton, Jonathan D R; Rangel, Drauzio E N; Cooke, Louise R; Hallsworth, John E

    2016-05-01

    There is a pressing need to understand and optimize biological control so as to avoid over-reliance on the synthetic chemical pesticides that can damage environmental and human health. This study focused on interactions between a novel biocontrol-strain, Bacillus sp. JC12GB43, and potato-pathogenic Phytophthora and Fusarium species. In assays carried out in vitro and on the potato tuber, the bacterium was capable of near-complete inhibition of pathogens. This Bacillus was sufficiently xerotolerant (water activity limit for growth = 0.928) to out-perform Phytophthora infestans (~0.960) and challenge Fusarium coeruleum (~0.847) and Fusarium sambucinum (~0.860) towards the lower limits of their growth windows. Under some conditions, however, strain JC12GB43 stimulated proliferation of the pathogens: for instance, Fusarium coeruleum growth-rate was increased under chaotropic conditions in vitro (132 mM urea) by >100% and on tubers (2-M glycerol) by up to 570%. Culture-based assays involving macromolecule-stabilizing (kosmotropic) compatible solutes provided proof-of-principle that the Bacillus may provide kosmotropic metabolites to the plant pathogen under conditions that destabilize macromolecular systems of the fungal cell. Whilst unprecedented, this finding is consistent with earlier reports that fungi can utilize metabolites derived from bacterial cells. Unless the antimicrobial activities of candidate biocontrol strains are assayed over a full range of field-relevant parameters, biocontrol agents may promote plant pathogen infections and thereby reduce crop yields. These findings indicate that biocontrol activity, therefore, ought to be regarded as a mode-of-behaviour (dependent on prevailing conditions) rather than an inherent property of a bacterial strain. PMID:26880001

  8. Phaeobacter inhibens as biocontrol agent against Vibrio vulnificus in oyster models

    DEFF Research Database (Denmark)

    Porsby, Cisse Hedegaard; Gram, Lone

    2016-01-01

    acid (TDA), inhibited V. vulnificus as did pure TDA (MIC of 1-3.9 μM). P. inhibens DSM 17395 (at 106 cfu/ml) eradicated 105 cfu/ml V. vulnificus CMCP6 (a rifampicin resistant variant) from a co-culture oyster model system (oyster juice) whereas the pathogen grew to 107 cfu/ml when co......, the presence of P. inhibens could not prevent subsequently added V. vulnificus from entering the live animals, likely because of too low levels of the biocontrol strain. Whilst the oyster model studies provided indication that P. inhibens DSM 17395 could be a good candidate as biocontrol agent against V...

  9. Colonization of three antagonistic strains on rice plant and their biocontrol effects on rice sheath blight%3株拮抗菌在水稻植株上的定殖能力及对纹枯病的防效

    Institute of Scientific and Technical Information of China (English)

    张德涛; 彭正凯; 曹琦琦; 杨媚; 郑丽; 周而勋

    2012-01-01

    【Objective】 The objective of this study was to explore the colonization ability and biocontrol effects of three antagonistic strains NB12(Bacillus sp.),SD7(Burkholderia sp.)and NB20(Streptomyces katrae)screened by our laboratory on rice sheath blight(RSB) at detached leave,seeding and tillering stage.【Method】 Mutants of strains NB12 and SD7 resistant to rifampin at 300 μg/mL were induced and screened at various concentrations of rifampin,and were used in their colonization study.RSB pathogen Rhizoctonia solani mycelial masses of diameter 5 mm were used as inoculum for inoculation on detached leave test,while rice grains covered with R.solani mycelia were used as inoculum for inoculation at seedling and tillering stages.【Result】 The results indicated that both strains of NB12 and SD7 could colonize on rice leaves at a high density.More than 106 CFU/g labeled strains could be detected after 20 days on mature leaves and labeled strains could be also detected after 10 days on newly-growing leaves.The control effects of strains NB12,SD7 and NB20 on RSB of detached leaves reached 91.3%,90.0% and 54.4%,respectively,significantly higher than that(77.5%) of Jinggangmycin at 40 μg/mL.The three strains showed promising control effect of 59.91%,53.73% and 38.47%,respectively,on RSB at seedling stage and their cell-free fermented filtrates also behaved certain effect.However,all of the three strains showed an unsatisfactory effect at tillering stage,and two better treatments of fermented fluids of NB12 and SD7 just behaved limited control effect of 17.6% and 24.0%,respectively.It was also found that the mixed fermented fluids of strain NB12,SD7 and NB20 could not only raise the control effect,but also lower it.【Conclusion】 Strains NB12 and SD7 behaved promising biocontrol ability to RSB at detached leave test and seedling stage,but the control effect on RSB at tillering stage needs further study.%【目的】研究

  10. Biocontrol of Soil Fungi in Tomato with Microencapsulates Containing Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Marcela H. Suarez

    2011-01-01

    Full Text Available Problem statement: An option to reduce pollution by synthetic agro-chemical in root plant disease management is the use of antagonist rhizobacteria belonging to Bacillus genus, because their inhibitory properties, stimulation of plant growth and crop yield increase. Approach: This study was carried out in order to evaluate if Bacillus subtilis strains could play an antagonists role of plant pathogens and if they can be microencapsulated inside a biopolymer matrix. It was adapted an equipment and evaluated a technique for microcapsules elaboration, in order to incorporate B. subtilis strains and to analyze their potential as biocontrol agents by determining their antagonistic effect against pathogenic soil fungi; in addition, it was analyzed their effect on tomato plant growth promotion under greenhouse conditions. B. subtilis strains identified as B1, J1, M2 and their mixture were used; microcapsules containing bacterial strains were inoculated to tomato seeds cv. Floradade. When seedlings emerged, a second application of microcapsules containing B. subtilis was performed on the pots, which previously were inoculated with the fungi Rhizoctonia solani and Fusarium oxysporum. Response variables were: Incidence and disease severity, plant growth, aerial and root dry weight, leaf area and fruit yield. Results: The outcome showed that the equipment designed and adapted for microcapsules elaboration was useful to obtain microcapsules containing the bacterial strains. B. subtilis strains exerted apparent biocontrol, since incidence and disease severity was reduced and for that reason inhibited the infective activity of the inoculated plant pathogens, also microcapsules containing Bacillus strains stimulated tomato growth and fruit yield. Conclusion: Microcapsules containing B. subtilis strains could be effective biocontrol agents against soil fungi plant pathogens and could have a potential biofertilizer effect, since they stimulated growth and yield

  11. Identification of Bio-Control Strain 1505 and Its Effect on Controlling Postharvest Citrus Anthracnose%生防细菌1505的鉴定及其对采后柑橘炭疽病的抑制效果

    Institute of Scientific and Technical Information of China (English)

    汪茜; 胡春锦; 黄思良; 柯仿钢

    2011-01-01

    采用细菌学方法及分子生物学技术对菌株1505进行鉴定,并对其在防治采后柑橘炭疽病上的效果进行研究.结果显示,菌株1505对柑橘炭疽病菌具有拮抗活性,其活菌悬浮液在PDA平板上对柑橘炭疽病菌产生明显抑制作用,而灭菌发酵液和去除菌体的发酵液均无抑菌效果;连续8次在人工培养基上转代培养,菌株1505对柑橘炭疽病菌生长的抑制力没有发生明显改变;菌株1505对胶孢炭疽菌[Colletotrichum gloeosporioides(Penz.)Sacc.]引起的柑橘炭疽病有明显的防治作用,刺伤接种的防效为48.8%,自然发病的防效为46.3%.根据形态学特征和生理生化反应,结合16SrDNA序列分析结果,将菌株1505鉴定为枯草芽孢杆菌(Bacillus subtilis).%The present study aimed to use the bacteriological method and molecular biology technique to identify the strain 1505 and study its efficacy in controlling postarvest Citrus anthracnose. The results showed that the strain 1505 had antagonistic actwity against Colletatrichum gloeosyorioides. its cell auspension showed significant suppressive activity against C. gloeosporioides , while no suppressive activity was detected in high.temperature-sterilized fermentation fluid or cell-free fermentation fluid. Lesa change in the suppressive ability of strain 1505 against C. gloeosporioides was observed during 8 continuously subculture in artificial media. The strain 1505 had significant efficacy in controlling Citrus anthracnose disease causing by C. gloeosporioides. The average efficacies of strain 1505 in controlling the disease were 48. 8 % in artificial inoculation test and 46. 3 % in natural infection test. Strain 1505 was identified as Bacillus subtilis based on the physiological and biochemical properties and its 16S rDNA sequence analysis.

  12. Commercialization of postharvest biocontrol: barriers and opportunities

    Science.gov (United States)

    The past twenty years has seen the field of postharvest biocontrol evolve from an unknown entity with one or two novel reports in the literature to a sophisticated science with strong research programs worldwide, hundreds of publications, patented technologies, and now several commercial products. ...

  13. Hrp mutant bacteria as biocontrol agents: toward a sustainable approach in the fight against plant pathogenic bacteria.

    Science.gov (United States)

    Hanemian, Mathieu; Zhou, Binbin; Deslandes, Laurent; Marco, Yves; Trémousaygue, Dominique

    2013-10-01

    Sustainable agriculture necessitates development of environmentally safe methods to protect plants against pathogens. Among these methods, application of biocontrol agents has been efficiently used to minimize disease development. Here we review current understanding of mechanisms involved in biocontrol of the main Gram-phytopathogenic bacteria-induced diseases by plant inoculation with strains mutated in hrp (hypersensitive response and pathogenicity) genes. These mutants are able to penetrate plant tissues and to stimulate basal resistance of plants. Novel protection mechanisms involving the phytohormone abscisic acid appear to play key roles in the biocontrol of wilt disease induced by Ralstonia solanacearum in Arabidopsis thaliana. Fully understanding these mechanisms and extending the studies to other pathosystems are still required to evaluate their importance in disease protection.

  14. 冬凌草内生细菌的分离鉴定及其对植物病害的生防作用%Isolation & Identification of Entophytic Bacterial Strain from Rabdosia rubescens & Its Biocontrol Effects against Plant Pathogens

    Institute of Scientific and Technical Information of China (English)

    柯杨; 马瑜; 沈莹华; 李勃

    2013-01-01

    An enlophylic bacterial strain KD3 isolated from the tissue of Rabdosia rubescens ( Hemsl. ) H. Hara obviously had antagonism against many crop fungal pathogens. This strain was identified as Bacillus sublilis according to its morphological, physiological and biochemical characteristics and 16S rRNA sequence analysis. The results of antagonism test showed that strain KD3 could control the plant pathogens effectively, which exhibited a good application and development potential.%从冬凌草(Rabdosia rubescens (Hemsl.) H.Hara)中分离筛选出1株对多种作物真菌病害具有显著拮抗作用的细菌,命名为KD3.通过其形态特征和生理生化特性以及16S rRNA序列的同源性分析,鉴定该菌株为枯草芽胞杆菌(Bacillus subtilis).试验表明,KD3菌株能够显著抑制多种病原真菌的侵染,具有良好的应用开发潜力.

  15. 生防菌FM4B的鉴定及抗菌物质的性质研究%Identification on Biocontrol Bacillus Strain FM4B and Detection of Its Antagonistic Substance

    Institute of Scientific and Technical Information of China (English)

    胡雪芹; 芮广虎; 周雪梅; 张洪斌

    2011-01-01

    采用生理生化、16S rDNA等方法鉴定菌株FM4B并对该菌株抑菌物质粗提液的性质进行了初步研究.生理生化实验显示,菌株FM4B属于短短芽孢杆菌,16S rDNA基因的测定与分析表明,FM4B与B.brevis(短短芽孢杆菌)的同源性达到99.5%,故推定FM4B为短短芽孢杆菌.实验表明抑菌物质粗提物对多种细菌、真菌有抑制作用,并且耐高温(90℃)、耐酸碱(pH 2~11)、对蛋白酶K、胰蛋白酶、胃蛋白酶不敏感.%Classical physiological and biochemical methods and 16S rDNA analysis were used to identify bacillus strain FM4B and the crude extract produced by the strain was studied. Physiological and biochemical identification showed that FM4B related to Brevibacillus brevis. 16S rDNA analysis showed that the 16S rDNA sequence of FM4B shared 99.5% homologies with sequence of Brevibacillus brevis from GenBank. So it is considered that the strain of FM4B belongs to Brevibacillus brevis. The crude extract had antibacterial activity against bacteria and fungi, which was stable after heated (90℃) and pH altered (pH2 ~ 11), resistant to trypsin, proteinase K and pepsin.

  16. Isolation,Screening and Preliminary Identification of Biocontrol Strains of Bacillus spp.%生防芽孢杆菌的分离筛选与初步鉴定

    Institute of Scientific and Technical Information of China (English)

    任士伟; 邢小霞; 董向丽

    2011-01-01

    30 Strains of Bacillus spp. were isolated from 7 soil samples using repeated plate streaking, and 4 strains of them had rivalry ability to Fusarium oxysporum f. sp. cucumerinum. They had also antagonism effect and broad spectrum to other plant pathogenic fungi proved by the method of plate test. We detected that the secondary metabolites of Bacillus spp. had fungistasis and could induce deformed growth of hyphae, the metabolites had water-solubility ability. The hyphae of Fusarium oxysporum f. sp. cucumerinum et al. could be induced bulge, browning, tip dulled. produced sporule by zymotic fluid. A strain of Bacillus subtilis was preliminary identified through observation of its morphological features and culture characters and a series of biochemical experiments.%从不同环境中采集7份土壤样品用于芽孢杆菌的分离纯化,通过多次平板划线分离纯化得到30个菌落形态不同的菌株.以黄瓜枯萎病菌为指示菌对得到的菌株进行抑菌活性筛选,其中的4个菌株具有明显的抑菌作用.采用对峙生长法,测定4株芽孢杆菌对其它植物病原真菌的活性,结果显示4个芽孢杆菌均具有广谱抑菌性.进一步试验表明,4株芽孢杆菌的发酵液也具有明显的抑菌作用,说明对病原真菌有抑制作用的物质水溶性较大,大部分释放到发酵液中,并且发酵液可使黄瓜枯萎等植物病原真菌菌丝膨大、褐变、末端钝圆,促进镰刀菌产生小孢子.通过菌落形态特征、系列生化测定对其进行初步鉴定,结果表明其中一株为枯草芽孢杆菌.

  17. BSH-4菌株对黄瓜菌核病的防治作用及其鉴定%Biocontrol of bacterium strain BSH-4 against cucumber sclerotial blight and its identification

    Institute of Scientific and Technical Information of China (English)

    于婷; 潘金菊; 车亚莉; 慕卫; 刘峰

    2009-01-01

    An antagonistic bacterium strain BSH-4 was isolated from rhizosphere soil of cucumber in the greenhouse. Dual culture test indicated that the strain was highly inhibitory to Sclerotinia sclerotiorum by making the mycelium swollen and curly, the protoplasm inside brown and concentrated, and the production of sclerotium delayed. Treat with fermentation broth of BSH-4, the disease control efficacy was 81.6% on detached leaves test, and 68.3% on pot trials. And the recovering rate was attained to 74.4% when sclerotial blight infected stems of cucumber was treated in the field. All the efficacies of BSH-4 broth excelled that of dimethachlon fungicide. The fermentation broth of BSH-4 had no impact on seed germination rate, but also promoted the growth of cucumber seedling. Identification by morphological observation, physiological and biochemical determinations, and 16S rDNA sequences analysis showed that the strain was belonged to Bacillus pumilus.%从日光温室黄瓜根际土壤中分离获得1株拮抗细菌菌株BSH-4.采用平板对峙法结合显微镜观察确定BSH-4对黄瓜菌核病病菌具有较强的抑制作用,导致菌丝粗大、弯曲,菌丝体内原生质褐化、聚集,并延迟菌核形成的时间.BSH-4发酵液对离体叶片接种引起的黄瓜菌核病防效为81.6%,盆栽土壤接种防效为68.3%;对田间发病茎秆治愈率为74.4%,明显优于对照药剂菌核净,且对种子出苗率无影响,对瓜苗有促长作用.经形态特性观察,生理生化鉴定及16S rDNA序列分析,初步确定该菌株为短小芽孢杆菌Bacillus pumilus.

  18. SCREENING OF FLUORESCENT RHIZOBACTERIA FOR THE BIOCONTROL OF SOILBORNE PLANT PATHOGENIC FUNGI

    Directory of Open Access Journals (Sweden)

    ANELISE DIAS

    2014-01-01

    Full Text Available The biocontrol of soilborne plant pathogens represents a promising approach from the environ- mental and practical points of view. Fluorescent pseudomonad rhizobacteria are well known by their antagonis- tic capacity towards several plant pathogens due to a diversity of antimicrobial metabolites they produce. This study was conceived to select and characterize rhizobacteria having antagonistic potential towards the patho- genic fungi Rhizoctonia solani and Sclerotium rolfsii. A total of 94 bacterial strains isolated from the rhizospheres of four vegetable species under organic cultivation were evaluated. Twenty-two strains which predominate in lettuce and rudbeckia rhizospheres showed identical biochemical profiles to Pseudomonas fluo- rescens, while in kale and parsley rhizospheres identical profiles to Pseudomonas putida (subgroups A and B strains prevailed. Two types of antagonism were verified in vitro and defined as competition and inhibition of mycelial growth. Sixty percent of the evaluated strains showed antagonistic potential and, among those, 24 strains expressed antagonism to both target fungi, with P. fluorescens being the most representative bacterial species. This work clearly identified a number of strains with potential for use as plant growth-promoting and biocontrol of the two soilborne fungal pathogens in vegetable crops production systems.

  19. 荧光假单胞菌拮抗菌株对烟草疫霉的抑菌机制及控病效果%Antagonistic mechanism of Pseudomonas fluorescens strains against Phytophthora nicotianae and biocontrol effect on tobacco black shank

    Institute of Scientific and Technical Information of China (English)

    董国菊; 马冠华; 肖崇刚

    2012-01-01

    To explore the antagonistic mechanism of rhizospheric bacteria against Phytophthora nicotianae van Breda de Haan,five antagonistic Pseudomonas fluorescens strains were isolated from rhizospheric soils of tobacco fields in Chongqing.Efficient antagonistic strain against P.nicotianae was screened using dual culture technique and metabolites inhibitory test.Among these strains,P-72-10 showed the highest suppressive effect;it produced an inhibition zone of 13.0mm(radius) and an inhibitory rate at 68.57%.Furthermore,the growth of pathogen's hypha was also suppressed by the extracellular metabolites of strain P-72-10 from 25.39% to 46.03%.P.nicotianae was observed microscopically excessive branching of mycelia and malformation of mycelia tips;thick-walled cells with concentrated plasmas and chlamydospore-like cells.When tested on the tobacco plants grown in greenhouse,strain P-72-10 got a good biocontrol effect on tobacco black shank infection and effects on the resistant and susceptible varieties of tobacco were 53.57% and 66.37% respectively.%为探讨烟草根际生防细菌对烟草疫霉Phytophthora nicotianae的抑菌机制,从重庆地区连作烟田健康烟株根际土壤分离获得5株荧光假单胞菌Pseudomonas fluorescens拮抗菌株。通过平板对峙及代谢产物抑菌试验筛选对烟草疫霉具有高效拮抗作用的菌株,其中,P-72-10菌株抑菌效果最强,抑菌带半径达13.0 mm,相对抑制率为68.57%,且该菌株代谢产物对烟草疫霉菌丝生长有明显的抑制作用,相对抑制率达25.39%~46.03%;显微观察发现该菌株可引起烟草疫霉菌丝的分支增多,菌丝顶端膨大呈畸形,多数菌丝中间或顶端细胞的细胞壁加厚、原生质浓缩和产生类似厚壁孢子的细胞。在温室盆栽条件下P-72-10菌株对烟草黑胫病也表现出良好的控病效果,对抗病和感病品种的相对防效分别为53.57%和66.37%。

  20. Biocontrol effect and taxonomy of antagonistic Streptomyces strain B28%拮抗链霉菌B28的分类鉴定及其生防作用研究

    Institute of Scientific and Technical Information of China (English)

    申屠旭萍; 于冰; 边亚琳; 胡靓; 马正; 童朝明; 俞晓平

    2012-01-01

    目前植物真菌病害的防治仍以化学药剂为主.化学农药虽然具有毒力强、见效快等特点,但长期大量使用化学农药带来了环境污染、农药残留和病原菌的抗药性等诸多问题.因此,研制高效、低毒、和对环境安全的生物农药已迫在眉睫.放线菌是一类有着广泛实际用途的微生物资源,具有农用活性的放线菌是生物农药研制的重要出发菌株.世界上约有70%的抗生素是由放线菌代谢产生的,这些放线菌主要来源于土壤.%An antagonistic Streptomyces strain B28, isolated from the soil collected in Tianmu mountain of Zhejiang Province, was identified as Streptomyces diastatochromogenes by morphological , physiological & bi-ochemical characteristics and 16S rDNA sequence alignment. Antagonistic study indicated that its secondary metabolite toyocamycin had an antifungal activity on the mycelial growth of seven species of plant pathogenic fungi. The results showed that toyocamycin possessed marked inhibitory activity against Rhizoctonia solani and the EC50 value was 0.58 μg/mL. The effective of toyocamycin against Botrytis cinerea and Fusarium oxyspo-rum f. sp. cucumerinum was much higher than that against Colletotrichum gloeosporioides, the EC50, value of toyocamycin in inhibiting B. cinerea, F. oxysporum and C. gloeosporioides were 13.65 μg/mL, 12.33 μg/ mL, and 30.15 μg/mL respectively.

  1. Evaluation of economically feasible, natural plant extract-based microbiological media for producing biomass of the dry rot biocontrol strain Pseudomonas fluorescens P22Y05 in liquid culture.

    Science.gov (United States)

    Khalil, Sadia; Ali, Tasneem Adam; Skory, Chris; Slininger, Patricia J; Schisler, David A

    2016-02-01

    The production of microbial biomass in liquid media often represents an indispensable step in the research and development of bacterial and fungal strains. Costs of commercially prepared nutrient media or purified media components, however, can represent a significant hurdle to conducting research in locations where obtaining these products is difficult. A less expensive option for providing components essential to microbial growth in liquid culture is the use of extracts of fresh or dried plant products obtained by using hot water extraction techniques. A total of 13 plant extract-based media were prepared from a variety of plant fruits, pods or seeds of plant species including Allium cepa (red onion bulb), Phaseolus vulgaris (green bean pods), and Lens culinaris (lentil seeds). In shake flask tests, cell production by potato dry rot antagonist Pseudomonas fluorescens P22Y05 in plant extract-based media was generally statistically indistinguishable from that in commercially produced tryptic soy broth and nutrient broth as measured by optical density and colony forming units/ml produced (P ≤ 0.05, Fisher's protected LSD). The efficacy of biomass produced in the best plant extract-based media or commercial media was equivalent in reducing Fusarium dry rot by 50-96% compared to controls. In studies using a high-throughput microbioreactor, logarithmic growth of P22Y05 in plant extract-based media initiated in 3-5 h in most cases but specific growth rate and the time of maximum OD varied as did the maximum pH obtained in media. Nutrient analysis of selected media before and after cell growth indicated that nitrogen in the form of NH4 accumulated in culture supernatants, possibly due to unbalanced growth conditions brought on by a scarcity of simple sugars in the media tested. The potential of plant extract-based media to economically produce biomass of microbes active in reducing plant disease is considerable and deserves further research. PMID:26745985

  2. Evaluation of economically feasible, natural plant extract-based microbiological media for producing biomass of the dry rot biocontrol strain Pseudomonas fluorescens P22Y05 in liquid culture.

    Science.gov (United States)

    Khalil, Sadia; Ali, Tasneem Adam; Skory, Chris; Slininger, Patricia J; Schisler, David A

    2016-02-01

    The production of microbial biomass in liquid media often represents an indispensable step in the research and development of bacterial and fungal strains. Costs of commercially prepared nutrient media or purified media components, however, can represent a significant hurdle to conducting research in locations where obtaining these products is difficult. A less expensive option for providing components essential to microbial growth in liquid culture is the use of extracts of fresh or dried plant products obtained by using hot water extraction techniques. A total of 13 plant extract-based media were prepared from a variety of plant fruits, pods or seeds of plant species including Allium cepa (red onion bulb), Phaseolus vulgaris (green bean pods), and Lens culinaris (lentil seeds). In shake flask tests, cell production by potato dry rot antagonist Pseudomonas fluorescens P22Y05 in plant extract-based media was generally statistically indistinguishable from that in commercially produced tryptic soy broth and nutrient broth as measured by optical density and colony forming units/ml produced (P ≤ 0.05, Fisher's protected LSD). The efficacy of biomass produced in the best plant extract-based media or commercial media was equivalent in reducing Fusarium dry rot by 50-96% compared to controls. In studies using a high-throughput microbioreactor, logarithmic growth of P22Y05 in plant extract-based media initiated in 3-5 h in most cases but specific growth rate and the time of maximum OD varied as did the maximum pH obtained in media. Nutrient analysis of selected media before and after cell growth indicated that nitrogen in the form of NH4 accumulated in culture supernatants, possibly due to unbalanced growth conditions brought on by a scarcity of simple sugars in the media tested. The potential of plant extract-based media to economically produce biomass of microbes active in reducing plant disease is considerable and deserves further research.

  3. 黄麻链霉菌 NF0919菌株对草莓枯萎病的生防活性初探%A Primary Study on Biocontrol Activity of Streptomyces corchorusii Strain NF0919 to Strawberry Fusarium Wilt

    Institute of Scientific and Technical Information of China (English)

    陈宏州; 庄义庆; 杨敬辉

    2014-01-01

    采用菌丝生长速率法测定了黄麻链霉菌NF0919发酵液对草莓枯萎病菌的室内毒力,并进行了防治草莓枯萎病的室内盆栽试验。结果表明:NF0919菌株发酵液和申嗪霉素对草莓枯萎病菌菌丝生长抑制的EC50值分别为24.3752和0.4001μg/mL;NF0919菌株发酵上清20倍液和1%申嗪霉素SC 800倍液药后30 d对草莓枯萎病的防效分别为65%和90%。%We determined the indoor toxicity of the fermentation broth of Streptomyces corchorusii strain NF0919 to the pathogen ( Fusarium oxysporum f.sp.fragariae) of strawberry fusarium wilt through adopting mycelial growth rate method, and carried out the indoor pot experiment for the control of strawberry fusarium wilt.The results indicated that the EC50 values of NF0919 fermenta-tion broth and Phenazine-1-carboxylic against the mycelial growth of Fusarium oxysporum f.sp.fragariae were 24.3752 and 0.4001μg/mL respectively.On the 30th day after root-pouring treatment with the 20-fold dilution of fermentation supematant of NF0919 or the 800-fold dilution of 1%Phenazine-1-carboxylic SC, their control effect on strawberry fusarium wilt reached 65%or 90%, respectively.

  4. Genotypic comparison of Pantoea agglomerans plant and clinical strains

    OpenAIRE

    Frey Jürg E; Montesinos Emilio; Smits Theo HM; Rezzonico Fabio; Duffy Brion

    2009-01-01

    Abstract Background Pantoea agglomerans strains are among the most promising biocontrol agents for a variety of bacterial and fungal plant diseases, particularly fire blight of apple and pear. However, commercial registration of P. agglomerans biocontrol products is hampered because this species is currently listed as a biosafety level 2 (BL2) organism due to clinical reports as an opportunistic human pathogen. This study compares plant-origin and clinical strains in a search for discriminati...

  5. Screening of Trichoderma strains tolerant to benzimidazole

    Institute of Scientific and Technical Information of China (English)

    LIU Kai-qi; XIANG Mei-mei; LIU Ren; ZENG Yong-san; ZHOU Hong-zi; YU Jin-feng; JIANG Xin-yin; ZHANG Yue-li

    2004-01-01

    @@ The screening of isolates and the assay of biocontrol mechanisms of Trichoderma were studied systematically in laboratory and greenhouse in vivo. The proteins tolerant to benzimidazole in Trichoderma strains were purified, and their physical and chemical properties were detected. Compared their biological activities in vitro and vivo in greenhouse, nine biocontrol strains (including Ty- 10-2, LTR-2, Tj-5-1, Tj-5-4, Ty- 11-1, Tj-11-3, Ty- 11-3, Tj-3-3-2, Tj-3-3-4) were screened. These biocontrol strains had faster rates of growth and higher inhibition to gray mould (Bortrytis cinerea),and the inhibition was stable. The effects of controlling gray mould in greenhouse with the screened Trichoderma strains were 70 % and 50 % in vivo.

  6. Non-target effects of the microbial control agents Pseudomonas fluorescens DR54 and Clonostachys rosea IK726 in soils cropped with barley followed by sugar beet

    DEFF Research Database (Denmark)

    Johansen, Anders; Knudsen, Inge M.B.; Binnerup, Svend J.;

    2005-01-01

    Non-target effects of a bacterial (Pseudomonas fluorescens DR54) and a fungal (Clonostachys rosea IK726) microbial control agent (MCA), on the indigenous microbiota in bulk soil and rhizosphere of barley, and subsequent a sugar beet crop, were studied in a greenhouse experiment. MCAs were...... by a factor of 106 and 20, respectively, and DR54 showed a short-lasting growth increase in the sugar beet rhizosphere. In general, the non-target effects were small and transient. IK726 seemed to have general stimulating effects on soil enzyme activity and the soil microbiota, and resulted in a significant...... to be suitable for in situ monitoring of MCA non-target effects on the soil microbiota, but should be combined with assays for MCA survival and soil enzyme activity....

  7. ppGpp controlled by the Gac/Rsm regulatory pathway sustains biocontrol activity in Pseudomonas fluorescens CHA0.

    Science.gov (United States)

    Takeuchi, Kasumi; Yamada, Kosumi; Haas, Dieter

    2012-11-01

    In Pseudomonas fluorescens CHA0 and other fluorescent pseudomonads, the Gac/Rsm signal transduction pathway is instrumental for secondary metabolism and biocontrol of root pathogens via the expression of regulatory small RNAs (sRNAs). Furthermore, in strain CHA0, an imbalance in the Krebs cycle can affect the strain's ability to produce extracellular secondary metabolites, including biocontrol factors. Here, we report the metabolome of wild-type CHA0, a gacA-negative mutant, which has lost Gac/Rsm activities, and a retS-negative mutant, which shows strongly enhanced Gac/Rsm-dependent activities. Capillary electrophoresis-based metabolomic profiling revealed that the gacA and retS mutations had opposite effects on the intracellular levels of a number of central metabolites, suggesting that the Gac/Rsm pathway regulates not only secondary metabolism but also primary metabolism in strain CHA0. Among the regulated metabolites identified, the alarmone guanosine tetraphosphate (ppGpp) was characterized in detail by the construction of relA (for ppGpp synthase) and spoT (for ppGpp synthase/hydrolase) deletion mutants. In a relA spoT double mutant, ppGpp synthesis was completely abolished, the expression of Rsm sRNAs was attenuated, and physiological functions such as antibiotic production, root colonization, and plant protection were markedly diminished. Thus, ppGpp appears to be essential for sustaining epiphytic fitness and biocontrol activity of strain CHA0.

  8. Microencapsulation Technique with Organic Additives for Biocontrol Agents

    Directory of Open Access Journals (Sweden)

    Szczech Magdalena

    2016-06-01

    Full Text Available The aim of these studies was to develop an effective formulation for microbial strains active in plant protection. Emulsification technique in oil was used to produce alginate microcapsules supplemented with organic additives to improve production process and stability of the preparations during storage. The release of microorganisms from the capsules in soil and their effectiveness in biocontrol of Fusarium wilt were evaluated. Three bacterial strains Burkholderia cepacia strain CAT5, Bacillus spp. strains PZ9 and SZ61, and fungus Trichoderma virens TRS106 were immobilized separately in calcium alginate supplemented with chitosan, peat powder, or skim milk. The productivity of microcapsules was enhanced by 60% when peat was added to the alginate matrix. Peat reduced also contamination of the capsules during storage, significantly enhancing their quality. By contrast, the addition of skim milk reduced quality of the microcapsules. The additives did not influence the viability of entrapped microorganisms and their release in soil. The survival of the microbial cells was mainly related to the kind of microorganism used, and the highest viability showed Bacillus sp. PZ9 and Trichoderma TRS106. Lyophilization of the microcapsules appeared to be unfavorable by reducing microbial viability in the capsules and in the soil after application. The best properties: good storage ability and sufficient microbial release in the soil, exhibited wet microcapsules amended with peat. These capsules were used to control Fusarium wilt in tomato plants. The protective effect was obtained when the microcapsule-entrapped bacteria PZ9 were used. The effectiveness of this bacterium was comparable with fungicide. Peat-amended microcapsules entrapping Bacillus PZ9 showed the best quality and may have potential for commercial use.

  9. Lactic acid bacteria from fresh fruit and vegetables as biocontrol agents of phytopathogenic bacteria and fungi.

    Science.gov (United States)

    Trias, Rosalia; Bañeras, Lluís; Montesinos, Emilio; Badosa, Esther

    2008-12-01

    This study evaluated the efficacy of lactic acid bacteria (LAB) isolated from fresh fruits and vegetables as biocontrol agents against the phytopathogenic and spoilage bacteria and fungi, Xanthomonas campestris, Erwinia carotovora, Penicillium expansum, Monilinia laxa, and Botrytis cinerea. The antagonistic activity of 496 LAB strains was tested in vitro and all tested microorganisms except P. expansum were inhibited by at least one isolate. The 496 isolates were also analyzed for the inhibition of P. expansum infection in wounds of Golden Delicious apples. Four strains (TC97, AC318, TM319, and FF441) reduced the fungal rot diameter of the apples by 20%; only Weissella cibaria strain TM128 decreased infection levels by 50%. Cell-free supernatants of selected antagonistic bacteria were studied to determine the nature of the antimicrobial compounds produced. Organic acids were the preferred mediators of inhibition but hydrogen peroxide was also detected when strains BC48, TM128, PM141 and FF441 were tested against E. carotovora. While previous reports of antifungal activity by LAB are scarce, our results support the potential of LAB as biocontrol agents against postharvest rot. PMID:19204894

  10. Production of a newly isolated Paenibacillus polymyxa biocontrol agent using monosodium glutamate wastewater and potato wastewater.

    Science.gov (United States)

    Gu, Likun; Bai, Zhihui; Jin, Bo; Zhang, Jianyun; Li, Wenying; Zhuang, Guoqiang; Zhang, Hongxun

    2010-01-01

    A phyllosphere bacterial strain EBL-06 was isolated from wheat leaves. The morphology, cultural characteristics, phospholipid fatty acids, physiological and antagonistic fungus activities of this strain were investigated. A phylogenetic tree was constructed by comparing with the published 16S rDNA sequences of the relevant bacteria. The results showed that the isolate EBL-06 was a strain of Paenibacillus polymyxa; this strain performed a high level of antagonistic fungus activity toward a broad spectrum of phytopathogens, such as Botrytis cinerea, Cladosporium cucumerinum, Fusarium spp. The isolate EBL-06 can grow well using monosodium glutamate wastewater (MGW) and potato wastewater (PW) as culture medium. The maximum yield of 6.5 x 10(9) CFU/mL of the isolate EBL-06 anti-fungus biocontrol agent was reached in 15 hr cultivation at 28 degrees C, pH 6.0-7.5 using the mixture of MGW and PW (1:9). PMID:21174972

  11. Construction of engineering Trichoderma strains and their characteristics against tomato gray mold

    Institute of Scientific and Technical Information of China (English)

    LIU Xian; ZHUANG Jing-hua; GAO Zeng-gui; YANG Chang-cheng; CHEN Jie

    2004-01-01

    @@ The transformed Trichoderma strains Ttrm31, Ttrm34 and Ttrm55 were obtained from Trichoderma wild strain T21 mutated by REMI technique for more effective biocontrol of tomato gray mold (Botrytis cinerea) with Trichoderma agent. Those transformants appeared much better in biocontrol activity in vitro or in vivo against tomato gray mold were better than that of wild strain T21. The main results were as follow:

  12. Diversity, biocontrol, and plant growth promoting abilities of xylem residing bacteria from solanaceous crops.

    Science.gov (United States)

    Achari, Gauri A; Ramesh, Raman

    2014-01-01

    Eggplant (Solanum melongena L.) is one of the solanaceous crops of economic and cultural importance and is widely cultivated in the state of Goa, India. Eggplant cultivation is severely affected by bacterial wilt caused by Ralstonia solanacearum that colonizes the xylem tissue. In this study, 167 bacteria were isolated from the xylem of healthy eggplant, chilli, and Solanum torvum Sw. by vacuum infiltration and maceration. Amplified rDNA restriction analysis (ARDRA) grouped these xylem residing bacteria (XRB) into 38 haplotypes. Twenty-eight strains inhibited growth of R. solanacearum and produced volatile and diffusible antagonistic compounds and plant growth promoting substances in vitro. Antagonistic strains XB86, XB169, XB177, and XB200 recorded a biocontrol efficacy greater than 85% against BW and exhibited 12%-22 % increase in shoot length in eggplant in the greenhouse screening. 16S rRNA based identification revealed the presence of 23 different bacterial genera. XRB with high biocontrol and plant growth promoting activities were identified as strains of Staphylococcus sp., Bacillus sp., Streptomyces sp., Enterobacter sp., and Agrobacterium sp. This study is the first report on identity of bacteria from the xylem of solanaceous crops having traits useful in cultivation of eggplant. PMID:24963298

  13. Diversity, Biocontrol, and Plant Growth Promoting Abilities of Xylem Residing Bacteria from Solanaceous Crops

    Directory of Open Access Journals (Sweden)

    Gauri A. Achari

    2014-01-01

    Full Text Available Eggplant (Solanum melongena L. is one of the solanaceous crops of economic and cultural importance and is widely cultivated in the state of Goa, India. Eggplant cultivation is severely affected by bacterial wilt caused by Ralstonia solanacearum that colonizes the xylem tissue. In this study, 167 bacteria were isolated from the xylem of healthy eggplant, chilli, and Solanum torvum Sw. by vacuum infiltration and maceration. Amplified rDNA restriction analysis (ARDRA grouped these xylem residing bacteria (XRB into 38 haplotypes. Twenty-eight strains inhibited growth of R. solanacearum and produced volatile and diffusible antagonistic compounds and plant growth promoting substances in vitro. Antagonistic strains XB86, XB169, XB177, and XB200 recorded a biocontrol efficacy greater than 85% against BW and exhibited 12%–22 % increase in shoot length in eggplant in the greenhouse screening. 16S rRNA based identification revealed the presence of 23 different bacterial genera. XRB with high biocontrol and plant growth promoting activities were identified as strains of Staphylococcus sp., Bacillus sp., Streptomyces sp., Enterobacter sp., and Agrobacterium sp. This study is the first report on identity of bacteria from the xylem of solanaceous crops having traits useful in cultivation of eggplant.

  14. Genotypic comparison of Pantoea agglomerans plant and clinical strains

    Directory of Open Access Journals (Sweden)

    Frey Jürg E

    2009-09-01

    Full Text Available Abstract Background Pantoea agglomerans strains are among the most promising biocontrol agents for a variety of bacterial and fungal plant diseases, particularly fire blight of apple and pear. However, commercial registration of P. agglomerans biocontrol products is hampered because this species is currently listed as a biosafety level 2 (BL2 organism due to clinical reports as an opportunistic human pathogen. This study compares plant-origin and clinical strains in a search for discriminating genotypic/phenotypic markers using multi-locus phylogenetic analysis and fluorescent amplified fragment length polymorphisms (fAFLP fingerprinting. Results Majority of the clinical isolates from culture collections were found to be improperly designated as P. agglomerans after sequence analysis. The frequent taxonomic rearrangements underwent by the Enterobacter agglomerans/Erwinia herbicola complex may be a major problem in assessing clinical associations within P. agglomerans. In the P. agglomerans sensu stricto (in the stricter sense group, there was no discrete clustering of clinical/biocontrol strains and no marker was identified that was uniquely associated to clinical strains. A putative biocontrol-specific fAFLP marker was identified only in biocontrol strains. The partial ORF located in this band corresponded to an ABC transporter that was found in all P. agglomerans strains. Conclusion Taxonomic mischaracterization was identified as a major problem with P. agglomerans, and current techniques removed a majority of clinical strains from this species. Although clear discrimination between P. agglomerans plant and clinical strains was not obtained with phylogenetic analysis, a single marker characteristic of biocontrol strains was identified which may be of use in strain biosafety determinations. In addition, the lack of Koch's postulate fulfilment, rare retention of clinical strains for subsequent confirmation, and the polymicrobial nature of P

  15. Selection and characterization of Argentine isolates of Trichoderma harzianum for effective biocontrol of Septoria leaf blotch of wheat.

    Science.gov (United States)

    Stocco, Marina C; Mónaco, Cecilia I; Abramoff, Cecilia; Lampugnani, Gladys; Salerno, Graciela; Kripelz, Natalia; Cordo, Cristina A; Consolo, Verónica F

    2016-03-01

    Species of the genus Trichoderma are economically important as biocontrol agents, serving as a potential alternative to chemical control. The applicability of Trichoderma isolates to different ecozones will depend on the behavior of the strains selected from each zone. The present study was undertaken to isolate biocontrol populations of Trichoderma spp. from the Argentine wheat regions and to select and characterize the best strains of Trichoderma harzianum by means of molecular techniques. A total of 84 out of the 240 strains of Trichoderma were able to reduce the disease severity of the leaf blotch of wheat. Thirty-seven strains were selected for the reduction equal to or greater than 50% of the severity, compared with the control. The percentage values of reduction of the pycnidial coverage ranged between 45 and 80%. The same last strains were confirmed as T. harzianum by polymerase chain reaction amplification of internal transcribed spacers, followed by sequencing. Inter-simple sequence repeat was used to examine the genetic variability among isolates. This resulted in a total of 132 bands. Further numerical analysis revealed 19 haplotypes, grouped in three clusters (I, II, III). Shared strains, with different geographical origins and isolated in different years, were observed within each cluster. The origin of the isolates and the genetic group were partially related. All isolates from Paraná were in cluster I, all isolates from Lobería were in cluster II, and all isolates from Pergamino and Santa Fe were in cluster III. Our results suggest that the 37 native strains of T. harzianum are important in biocontrol programs and could be advantageous for the preparation of biopesticides adapted to the agroecological conditions of wheat culture.

  16. Selection and characterization of Argentine isolates of Trichoderma harzianum for effective biocontrol of Septoria leaf blotch of wheat.

    Science.gov (United States)

    Stocco, Marina C; Mónaco, Cecilia I; Abramoff, Cecilia; Lampugnani, Gladys; Salerno, Graciela; Kripelz, Natalia; Cordo, Cristina A; Consolo, Verónica F

    2016-03-01

    Species of the genus Trichoderma are economically important as biocontrol agents, serving as a potential alternative to chemical control. The applicability of Trichoderma isolates to different ecozones will depend on the behavior of the strains selected from each zone. The present study was undertaken to isolate biocontrol populations of Trichoderma spp. from the Argentine wheat regions and to select and characterize the best strains of Trichoderma harzianum by means of molecular techniques. A total of 84 out of the 240 strains of Trichoderma were able to reduce the disease severity of the leaf blotch of wheat. Thirty-seven strains were selected for the reduction equal to or greater than 50% of the severity, compared with the control. The percentage values of reduction of the pycnidial coverage ranged between 45 and 80%. The same last strains were confirmed as T. harzianum by polymerase chain reaction amplification of internal transcribed spacers, followed by sequencing. Inter-simple sequence repeat was used to examine the genetic variability among isolates. This resulted in a total of 132 bands. Further numerical analysis revealed 19 haplotypes, grouped in three clusters (I, II, III). Shared strains, with different geographical origins and isolated in different years, were observed within each cluster. The origin of the isolates and the genetic group were partially related. All isolates from Paraná were in cluster I, all isolates from Lobería were in cluster II, and all isolates from Pergamino and Santa Fe were in cluster III. Our results suggest that the 37 native strains of T. harzianum are important in biocontrol programs and could be advantageous for the preparation of biopesticides adapted to the agroecological conditions of wheat culture. PMID:26873560

  17. Isolation and identification of actinomycetes from a compost-amended soil with potential as biocontrol agents.

    Science.gov (United States)

    Cuesta, Gonzalo; García-de-la-Fuente, Rosana; Abad, Manuel; Fornes, Fernando

    2012-03-01

    The search for new biocontrol strategies to inhibit the growth of phytopathogenic microorganisms has become widely widespread due to environmental concerns. Among actinomycetes, Streptomyces species have been extensively studied since they have been recognized as important sources of antibiotics. Actinomycete strains were isolated from a calcareous soil, 2 two-phase olive mill waste ('alperujo') composts, and the compost-amended soil by using selective media, and they were then co-cultured with 5 phytopathogenic fungi and 1 bacterium to perform an in vitro antagonism assay. Forty-nine actinomycete strains were isolated, 12 of them showing a great antagonistic activity towards the phytopathogenic microorganisms tested. Isolated strains were identified by 16S rDNA sequence analysis and phenotypic procedures. Eleven isolates concerned the genus Streptomyces and 1 actinomycete with chitinolytic activity belonged to the genus Lechevalieria. PMID:21190787

  18. Role of gluconic acid production in the regulation of biocontrol traits of Pseudomonas fluorescens CHA0.

    Science.gov (United States)

    de Werra, Patrice; Péchy-Tarr, Maria; Keel, Christoph; Maurhofer, Monika

    2009-06-01

    The rhizobacterium Pseudomonas fluorescens CHA0 promotes the growth of various crop plants and protects them against root diseases caused by pathogenic fungi. The main mechanism of disease suppression by this strain is the production of the antifungal compounds 2,4-diacetylphloroglucinol (DAPG) and pyoluteorin (PLT). Direct plant growth promotion can be achieved through solubilization of inorganic phosphates by the production of organic acids, mainly gluconic acid, which is one of the principal acids produced by Pseudomonas spp. The aim of this study was to elucidate the role of gluconic acid production in CHA0. Therefore, mutants were created with deletions in the genes encoding glucose dehydrogenase (gcd) and gluconate dehydrogenase (gad), required for the conversion of glucose to gluconic acid and gluconic acid to 2-ketogluconate, respectively. These enzymes should be of predominant importance for rhizosphere-colonizing biocontrol bacteria, as major carbon sources provided by plant root exudates are made up of glucose. Our results show that the ability of strain CHA0 to acidify its environment and to solubilize mineral phosphate is strongly dependent on its ability to produce gluconic acid. Moreover, we provide evidence that the formation of gluconic acid by CHA0 completely inhibits the production of PLT and partially inhibits that of DAPG. In the Deltagcd mutant, which does not produce gluconic acid, the enhanced production of antifungal compounds was associated with improved biocontrol activity against take-all disease of wheat, caused by Gaeumannomyces graminis var. tritici. This study provides new evidence for a close association of gluconic acid metabolism with antifungal compound production and biocontrol activity in P. fluorescens CHA0.

  19. Bio-Control of Salmonella Enteritidis in Foods Using Bacteriophages

    Directory of Open Access Journals (Sweden)

    Hongduo Bao

    2015-08-01

    Full Text Available Two lytic phages, vB_SenM-PA13076 (PA13076 and vB_SenM-PC2184 (PC2184, were isolated from chicken sewage and characterized with host strains Salmonella Enteritidis (SE ATCC13076 and CVCC2184, respectively. Transmission electron microscopy revealed that they belonged to the family Myoviridae. The lytic abilities of these two phages in liquid culture showed 104 multiplicity of infection (MOI was the best in inhibiting bacteria, with PC2184 exhibiting more activity than PA13076. The two phages exhibited broad host range within the genus Salmonella. Phage PA13076 and PC2184 had a lytic effect on 222 (71.4% and 298 (95.8% of the 311 epidemic Salmonella isolates, respectively. We tested the effectiveness of phage PA13076 and PC2184 as well as a cocktail combination of both in three different foods (chicken breast, pasteurized whole milk and Chinese cabbage contaminated with SE. Samples were spiked with 1 × 104 CFU individual SE or a mixture of strains (ATCC13076 and CVCC2184, then treated with 1 × 108 PFU individual phage or a two phage cocktail, and incubated at 4 °C or 25 °C for 5 h. In general, the inhibitory effect of phage and phage cocktail was better at 4 °C than that at 25 °C, whereas the opposite result was observed in Chinese cabbage, and phage cocktail was better than either single phage. A significant reduction in bacterial numbers (1.5–4 log CFU/sample, p < 0.05 was observed in all tested foods. The two phages on the three food samples were relatively stable, especially at 4 ºC, with the phages exhibiting the greatest stability in milk. Our research shows that our phages have potential effectiveness as a bio-control agent of Salmonella in foods.

  20. Bio-Control of Salmonella Enteritidis in Foods Using Bacteriophages

    Science.gov (United States)

    Bao, Hongduo; Zhang, Pengyu; Zhang, Hui; Zhou, Yan; Zhang, Lili; Wang, Ran

    2015-01-01

    Two lytic phages, vB_SenM-PA13076 (PA13076) and vB_SenM-PC2184 (PC2184), were isolated from chicken sewage and characterized with host strains Salmonella Enteritidis (SE) ATCC13076 and CVCC2184, respectively. Transmission electron microscopy revealed that they belonged to the family Myoviridae. The lytic abilities of these two phages in liquid culture showed 104 multiplicity of infection (MOI) was the best in inhibiting bacteria, with PC2184 exhibiting more activity than PA13076. The two phages exhibited broad host range within the genus Salmonella. Phage PA13076 and PC2184 had a lytic effect on 222 (71.4%) and 298 (95.8%) of the 311 epidemic Salmonella isolates, respectively. We tested the effectiveness of phage PA13076 and PC2184 as well as a cocktail combination of both in three different foods (chicken breast, pasteurized whole milk and Chinese cabbage) contaminated with SE. Samples were spiked with 1 × 104 CFU individual SE or a mixture of strains (ATCC13076 and CVCC2184), then treated with 1 × 108 PFU individual phage or a two phage cocktail, and incubated at 4 °C or 25 °C for 5 h. In general, the inhibitory effect of phage and phage cocktail was better at 4 °C than that at 25 °C, whereas the opposite result was observed in Chinese cabbage, and phage cocktail was better than either single phage. A significant reduction in bacterial numbers (1.5–4 log CFU/sample, p < 0.05) was observed in all tested foods. The two phages on the three food samples were relatively stable, especially at 4 °C, with the phages exhibiting the greatest stability in milk. Our research shows that our phages have potential effectiveness as a bio-control agent of Salmonella in foods. PMID:26305252

  1. Antagonistic and Biocontrol Potential of Trichoderma asperellum ZJSX5003 Against the Maize Stalk Rot Pathogen Fusarium graminearum.

    Science.gov (United States)

    Li, Yaqian; Sun, Ruiyan; Yu, Jia; Saravanakumar, Kandasamy; Chen, Jie

    2016-09-01

    The efficacy of seven strains of Trichoderma asperellum collected from the fields in Southern China was assessed against Fusarium graminearum (FG) the causal agent of corn stalk rot of maize were in vitro for their antagonistic properties followed by statistical model of principal compound analysis to identify the beneficial antagonist T. asperellum strain. The key factors of antagonist activity were attributed to a total of 13 factors including cell wall degrading enzymes (chitnase, protease and β-glucanases), secondary metabolites and peptaibols and these were analyzed from eight strains of Trichoderma. A linear regression model demonstrated that interaction of enzymes and secondary metabolites of T. asperellum strain ZJSX5003 enhanced the antagonist activity against FG. Further, this strain displayed a disease reduction of 71 % in maize plants inoculated with FG compared to negative control. Pointing out that the T. asperellum strain ZJSX5003 is a potential source for the development of a biocontrol agent against corn stalk rot.

  2. Antagonistic and Biocontrol Potential of Trichoderma asperellum ZJSX5003 Against the Maize Stalk Rot Pathogen Fusarium graminearum.

    Science.gov (United States)

    Li, Yaqian; Sun, Ruiyan; Yu, Jia; Saravanakumar, Kandasamy; Chen, Jie

    2016-09-01

    The efficacy of seven strains of Trichoderma asperellum collected from the fields in Southern China was assessed against Fusarium graminearum (FG) the causal agent of corn stalk rot of maize were in vitro for their antagonistic properties followed by statistical model of principal compound analysis to identify the beneficial antagonist T. asperellum strain. The key factors of antagonist activity were attributed to a total of 13 factors including cell wall degrading enzymes (chitnase, protease and β-glucanases), secondary metabolites and peptaibols and these were analyzed from eight strains of Trichoderma. A linear regression model demonstrated that interaction of enzymes and secondary metabolites of T. asperellum strain ZJSX5003 enhanced the antagonist activity against FG. Further, this strain displayed a disease reduction of 71 % in maize plants inoculated with FG compared to negative control. Pointing out that the T. asperellum strain ZJSX5003 is a potential source for the development of a biocontrol agent against corn stalk rot. PMID:27407296

  3. Selection rhizosphere-competent microbes for development of microbial products as biocontrol agents

    Science.gov (United States)

    Mashinistova, A. V.; Elchin, A. A.; Gorbunova, N. V.; Muratov, V. S.; Kydralieva, K. A.; Khudaibergenova, B. M.; Shabaev, V. P.; Jorobekova, Sh. J.

    2009-04-01

    Rhizosphere-borne microorganisms reintroduced to the soil-root interface can establish without inducing permanent disturbance in the microbial balance and effectively colonise the rhizosphere due to carbon sources of plant root exudates. A challenge for future development of microbial products for use in agriculture will be selection of rhizosphere-competent microbes that both protect the plant from pathogens and improve crop establishment and persistence. In this study screening, collection, identification and expression of stable and technological microbial strains living in soils and in the rhizosphere of abundant weed - couch-grass Elytrigia repens L. Nevski were conducted. A total of 98 bacteria isolated from the rhizosphere were assessed for biocontrol activity in vitro against phytopathogenic fungi including Fusarium culmorum, Fusarium heterosporum, Fusarium oxysporum, Drechslera teres, Bipolaris sorokiniana, Piricularia oryzae, Botrytis cinerea, Colletothrichum atramentarium and Cladosporium sp., Stagonospora nodorum. Biocontrol activity were performed by the following methods: radial and parallel streaks, "host - pathogen" on the cuts of wheat leaves. A culture collection comprising 64 potential biocontrol agents (BCA) against wheat and barley root diseases has been established. Of these, the most effective were 8 isolates inhibitory to at least 4 out of 5 phytopathogenic fungi tested. The remaining isolates inhibited at least 1 of 5 fungi tested. Growth stimulating activity of proposed rhizobacteria-based preparations was estimated using seedling and vegetative pot techniques. Seeds-inoculation and the tests in laboratory and field conditions were conducted for different agricultural crops - wheat and barley. Intact cells, liquid culture filtrates and crude extracts of the four beneficial bacterial strains isolated from the rhizosphere of weed were studied to stimulate plant growth. As a result, four bacterial strains selected from rhizosphere of weed

  4. Selection rhizosphere-competent microbes for development of microbial products as biocontrol agents

    Science.gov (United States)

    Mashinistova, A. V.; Elchin, A. A.; Gorbunova, N. V.; Muratov, V. S.; Kydralieva, K. A.; Khudaibergenova, B. M.; Shabaev, V. P.; Jorobekova, Sh. J.

    2009-04-01

    Rhizosphere-borne microorganisms reintroduced to the soil-root interface can establish without inducing permanent disturbance in the microbial balance and effectively colonise the rhizosphere due to carbon sources of plant root exudates. A challenge for future development of microbial products for use in agriculture will be selection of rhizosphere-competent microbes that both protect the plant from pathogens and improve crop establishment and persistence. In this study screening, collection, identification and expression of stable and technological microbial strains living in soils and in the rhizosphere of abundant weed - couch-grass Elytrigia repens L. Nevski were conducted. A total of 98 bacteria isolated from the rhizosphere were assessed for biocontrol activity in vitro against phytopathogenic fungi including Fusarium culmorum, Fusarium heterosporum, Fusarium oxysporum, Drechslera teres, Bipolaris sorokiniana, Piricularia oryzae, Botrytis cinerea, Colletothrichum atramentarium and Cladosporium sp., Stagonospora nodorum. Biocontrol activity were performed by the following methods: radial and parallel streaks, "host - pathogen" on the cuts of wheat leaves. A culture collection comprising 64 potential biocontrol agents (BCA) against wheat and barley root diseases has been established. Of these, the most effective were 8 isolates inhibitory to at least 4 out of 5 phytopathogenic fungi tested. The remaining isolates inhibited at least 1 of 5 fungi tested. Growth stimulating activity of proposed rhizobacteria-based preparations was estimated using seedling and vegetative pot techniques. Seeds-inoculation and the tests in laboratory and field conditions were conducted for different agricultural crops - wheat and barley. Intact cells, liquid culture filtrates and crude extracts of the four beneficial bacterial strains isolated from the rhizosphere of weed were studied to stimulate plant growth. As a result, four bacterial strains selected from rhizosphere of weed

  5. Enhanced biocontrol activity of Trichoderma through inactivation of a mitogen-activated protein kinase.

    Science.gov (United States)

    Mendoza-Mendoza, Artemio; Pozo, María J; Grzegorski, Darlene; Martínez, Pedro; García, Juan M; Olmedo-Monfil, Vianey; Cortés, Carlos; Kenerley, Charles; Herrera-Estrella, Alfredo

    2003-12-23

    The production of lytic enzymes in Trichoderma is considered determinant in its parasitic response against fungal species. A mitogen-activated protein kinase encoding gene, tvk1, from Trichoderma virens was cloned, and its role during the mycoparasitism, conidiation, and biocontrol was examined in tvk1 null mutants. These mutants showed a clear increase in the level of the expression of mycoparasitism-related genes under simulated mycoparasitism and during direct confrontation with the plant pathogen Rhizoctonia solani. The null mutants displayed an increased protein secretion phenotype as measured by the production of lytic enzymes in culture supernatant compared to the wild type. Consistently, biocontrol assays demonstrated that the null mutants were considerably more effective in disease control than the wild-type strain or a chemical fungicide. In addition, tvk1 gene disruptant strains sporulated abundantly in submerged cultures, a condition that is not conducive to sporulation in the wild type. These data suggest that Tvk1 acts as a negative modulator during host sensing and sporulation in T. virens. PMID:14673101

  6. Combining fluorescent Pseudomonas spp. strains to enhance suppression of fusarium wilt of radish

    OpenAIRE

    de Boer, Marjan; Sluis, Ientse van der; Loon, L.C. van; Bakker, P. A. H. M.

    2001-01-01

    Fusarium wilt diseases, caused by the fungus Fusarium oxysporum, lead to significant yield losses of crops. One strategy to control fusarium wilt is the use of antagonistic, root-colonizing Pseudomonas spp. It has been demonstrated that different strains of these bacteria suppress disease by different mechanisms. Therefore, application of a mixture of these biocontrol strains, and thus of several suppressive mechanisms, may represent a viable control strategy. A prerequisite for biocontrol by...

  7. Survival of the biocontrol agents Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116 on the spikes of barley in the field

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xin; ZHANG Bing-xin; ZHANG Zhen; SHEN Wei-feng; YANG Ching-hong; YU Jing-quan; ZHAO Yu-hua

    2005-01-01

    Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating disease that results in extensive yield losses to wheat and barley. A green fluorescent protein (GFP) expressing plasmid pRP22-GFP was constructed for monitoring the colonization of two biocontrol agents, Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116, on the spikes of barley and their effect on suppression of FHB. Survival and colonization of the Brevibacillus brevis ZJY- 1 and Bacillus subtilis ZJY- 116 strains on spikes of barley were observed by tracking the bacterial transformants with GFP expression. Our field study revealed that plasmid pRP22-GFP was stably maintained in the bacterial strains without selective pressure. The retrieved GFP-tagged strains showed that the bacterial population fluctuation accorded with that of the rain events. Furthermore, both biocontrol strains gave significant protection against FHB on spikes of barley in fields. The greater suppression of barley FHB disease was resulted from the treatment of barley spikes with biocontrol agents before inoculation with F. graminearum.

  8. Potential biocontrol agents for biofouling on artificial structures.

    Science.gov (United States)

    Atalah, Javier; Newcombe, Emma M; Hopkins, Grant A; Forrest, Barrie M

    2014-09-01

    The accumulation of biofouling on coastal structures can lead to operational impacts and may harbour problematic organisms, including non-indigenous species. Benthic predators and grazers that can supress biofouling, and which are able to be artificially enhanced, have potential value as augmentative biocontrol agents. The ability of New Zealand native invertebrates to control biofouling on marina pontoons and wharf piles was tested. Caging experiments evaluated the ability of biocontrol to mitigate established biofouling, and to prevent fouling accumulation on defouled surfaces. On pontoons, the gastropods Haliotis iris and Cookia sulcata reduced established biofouling cover by >55% and largely prevented the accumulation of new biofouling over three months. On wharf piles C. sulcata removed 65% of biofouling biomass and reduced its cover by 73%. C. sulcata also had better retention and survival rates than other agents. Augmentative biocontrol has the potential to be an effective method to mitigate biofouling on marine structures.

  9. Identification of bio-control bacterial strain KJB04-11 against soybean root rot disease and the produced lipopeptide antibiotics by the strain%大豆根腐病生防菌KJB04-11的鉴定及其产生的脂肽类抗生素

    Institute of Scientific and Technical Information of China (English)

    李旭; 马福海; 王秀娟; 孙健; 张惠文

    2012-01-01

    从大豆根围筛选到1株对尖孢镰刀菌和立枯丝核菌都具有很好拮抗作用的菌株KJB04-11,经形态观察、生理生化特征和16S rDNA序列分析,属于枯草芽孢杆菌(Bacillussubtilis).具有抗菌活性的KJB04-11发酵液无菌滤液对热和酸碱具有较强的稳定性.采用Sephadex G-25柱层析、反相HPLC和冷冻干燥从KJB04-11发酵液中分离纯化了抗菌活性成分.由红外光谱、MALDI-TOF-MS、氨基酸组成及脂肽合成酶基因扩增结果推测该菌株产生的抗菌物质为C16、C17的mycosubtilin和C15的surfactin.田间试验表明,大豆种子经KJB04-11发酵液包衣处理对大豆根腐病防效为53.6%,大豆产量提高12.5%.%A bacterial strain KJB04-11 with antagonistic effect against Fusarium oxysporum and Rhizoctonia solani was isolated from soybean rhizosphere. The strain was identified as Bacillus subtilis, based on its morphological, cultural, physiological, and biochemical characteristics, and 16S rDNA sequence analysis. The fermented filtrate with antagonistic activity showed certain thermostability and certain acidic and alkaline stability in the range of pH 1.0-13.0. The antagonistic active compositions in the fermented filtrate were purified by gel filtration on Sephadex G-25 column and RF-HPLC, and preliminarily identified as Cl6 mycosubtilin, C17 mycosubtilin, and C15 surfactin by infrared spectra, MALDI-TOF mass spectrum, amino acid composition analysis, and amplification of lipopeptide synthase gene. In field trial, the control efficacy of seed coating agent prepared with KJB04-11 against soybean root rot disease was 53. 6% , and the soybean yield was increased by 12.5% .

  10. Screening of Mutation High-Yielding Biocontrol Bacterium BJ1 by Ion Beam Irradiation and Effect of Controlling Fusarium oxysporum cucunerinum Disease

    International Nuclear Information System (INIS)

    BJ1 of Bacillus subtilis is an important biocontrol factor in control of fungus disease. In order to improve the antagonistic ability of the strain,and obtain high-efficiency strains, 12C6+ of different doses and linear energy transfer (LET) was used to irradiate the biocontrol bacterium BJ1. The optimum dose and LET of ion beam irradiation for the BJ1 are 200-400 Gy and 60 keV/μm,respectively. The antagonistic ability is increased by 2%-21%. The control effect of mutation to Fusarium oxysporum f. sp. cucunerinum is increased by 17.48% over that of BJ1, and mutation also has better plant growth-promoting effect. (authors)

  11. Phytohormone profiles induced by trichoderma isolates correspond with their biocontrol and plant growth-promoting activity on melon plants.

    Science.gov (United States)

    Martínez-Medina, Ainhoa; Del Mar Alguacil, Maria; Pascual, Jose A; Van Wees, Saskia C M

    2014-07-01

    The application of Trichoderma strains with biocontrol and plant growth-promoting capacities to plant substrates can help reduce the input of chemical pesticides and fertilizers in agriculture. Some Trichoderma isolates can directly affect plant pathogens, but they also are known to influence the phytohormonal network of their host plant, thus leading to an improvement of plant growth and stress tolerance. In this study, we tested whether alterations in the phytohormone signature induced by different Trichoderma isolates correspond with their ability for biocontrol and growth promotion. Four Trichoderma isolates were collected from agricultural soils and were identified as the species Trichoderma harzianum (two isolates), Trichoderma ghanense, and Trichoderma hamatum. Their antagonistic activity against the plant pathogen Fusarium oxysporum f. sp. melonis was tested in vitro, and their plant growth-promoting and biocontrol activity against Fusarium wilt on melon plants was examined in vivo, and compared to that of the commercial strain T. harzianum T-22. Several growth- and defense-related phytohormones were analyzed in the shoots of plants that were root-colonized by the different Trichoderma isolates. An increase in auxin and a decrease in cytokinins and abscisic acid content were induced by the isolates that promoted the plant growth. Principal component analysis (PCA) was used to evaluate the relationship between the plant phenotypic and hormonal variables. PCA pointed to a strong association of auxin induction with plant growth stimulation by Trichoderma. Furthermore, the disease-protectant ability of the Trichoderma strains against F. oxysporum infection seems to be more related to their induced alterations in the content of the hormones abscisic acid, ethylene, and the cytokinin trans-zeatin riboside than to the in vitro antagonism activity against F. oxysporum.

  12. Challenges and opportunities for the commercialization of postharvest biocontrol

    Science.gov (United States)

    The past twenty years has seen the field of postharvest biocontrol evolve into a sophisticated science with global research programs worldwide, numerous yearly publications, patented technologies, and the development of new commercial products. The use of these products, however, still remains limi...

  13. Screening of biocontrol agents for control of foliar diseases

    NARCIS (Netherlands)

    Köhl, J.

    2009-01-01

    Candidate antagonists for the development of biocontrol agents have to fulfill many criteria. The criterion often investigated first in detail is the antagonistic potential of candidates against the target pathogen. However, candidates must also have high ecological competence, must be suitable for

  14. Grouper as a natural biocontrol of invasive lionfish.

    Science.gov (United States)

    Mumby, Peter J; Harborne, Alastair R; Brumbaugh, Daniel R

    2011-01-01

    Lionfish (Pterois volitans/miles) have invaded the majority of the Caribbean region within five years. As voracious predators of native fishes with a broad habitat distribution, lionfish are poised to cause an unprecedented disruption to coral reef diversity and function. Controls of lionfish densities within its native range are poorly understood, but they have been recorded in the stomachs of large-bodied Caribbean groupers. Whether grouper predation of lionfish is sufficient to act as a biocontrol of the invasive species is unknown, but pest biocontrol by predatory fishes has been reported in other ecosystems. Groupers were surveyed along a chain of Bahamian reefs, including one of the region's most successful marine reserves which supports the top one percentile of Caribbean grouper biomass. Lionfish biomass exhibited a 7-fold and non-linear reduction in relation to the biomass of grouper. While Caribbean grouper appear to be a biocontrol of invasive lionfish, the overexploitation of their populations by fishers, means that their median biomass on Caribbean reefs is an order of magnitude less than in our study. Thus, chronic overfishing will probably prevent natural biocontrol of lionfishes in the Caribbean. PMID:21731769

  15. Grouper as a natural biocontrol of invasive lionfish.

    Directory of Open Access Journals (Sweden)

    Peter J Mumby

    Full Text Available Lionfish (Pterois volitans/miles have invaded the majority of the Caribbean region within five years. As voracious predators of native fishes with a broad habitat distribution, lionfish are poised to cause an unprecedented disruption to coral reef diversity and function. Controls of lionfish densities within its native range are poorly understood, but they have been recorded in the stomachs of large-bodied Caribbean groupers. Whether grouper predation of lionfish is sufficient to act as a biocontrol of the invasive species is unknown, but pest biocontrol by predatory fishes has been reported in other ecosystems. Groupers were surveyed along a chain of Bahamian reefs, including one of the region's most successful marine reserves which supports the top one percentile of Caribbean grouper biomass. Lionfish biomass exhibited a 7-fold and non-linear reduction in relation to the biomass of grouper. While Caribbean grouper appear to be a biocontrol of invasive lionfish, the overexploitation of their populations by fishers, means that their median biomass on Caribbean reefs is an order of magnitude less than in our study. Thus, chronic overfishing will probably prevent natural biocontrol of lionfishes in the Caribbean.

  16. Antifungal Activity of Isolated Bacillus amyloliquefaciens SYBC H47 for the Biocontrol of Peach Gummosis

    Science.gov (United States)

    Zhang, Yanzhou; Wei, Zhiwen; Guan, Zhengbing; Cai, Yujie; Liao, Xiangru

    2016-01-01

    The gummosis disease is caused by Botryosphaeria dothidea (Moug. ex. Fr) Ces. et de Not., and it is one of the most important diseases of stone fruits worldwide. The use of biocontrol as an alternative approach to synthetic chemical fungicides has aroused general concern about how to control plant diseases that are caused by phytopathogens. The aim of this study is to isolate Bacillus strains from raw honeys with the capacity to inhibit B. dothidea and to explore the mechanisms by which they could be used in the biocontrol of peach gummosis. Bacillus amyloliquefaciens SYBC H47 was isolated and identified on the basis of its physiological and biochemical characteristics and its 16S rRNA and gyrB gene sequences. The cell suspension and the cell-free supernatant of its culture showed significant antifungal activity against Aspergillus niger, Mucor racemosus, Fusarium oxysporum, Penicillium citrinum, and Candida albicans by agar-diffusion assays. The primary antifungal substances were bacillomycin L, fengycin, and surfactin, which were analyzed by HPLC LC/ESI-MS/MS. Bacillomycin L showed the best inhibitory effect against conidial germination of B. dothidea, followed by fengycin and surfactin. Surfactin had limited effects on mycelial growth, contrary to those of bacillomycin L and fengycin. However, a mixture of the three lipopeptides had a synergistic effect that disrupted the structure of the conidia and mycelia. In order to reduce the production cost, the use of waste frying peanut oil and soy oil as the sole carbon source increased the lipopeptide yield levels by approximately 17% (2.42 g/L) and 110% (4.35 g/L), respectively. In a field trial, the decreases in the infected gummosis rate (IGR) and the disease severity index (DSI) through cell suspension treatments were 20% and 57.5% (in 2014), respectively, and 40% and 57.5% (in 2015), respectively, in comparison with the control. In conclusion, B. amyloliquefaciens SYBC H47 could inhibit the germination of conidia

  17. Antifungal Activity of Isolated Bacillus amyloliquefaciens SYBC H47 for the Biocontrol of Peach Gummosis.

    Science.gov (United States)

    Li, Xunhang; Zhang, Yanzhou; Wei, Zhiwen; Guan, Zhengbing; Cai, Yujie; Liao, Xiangru

    2016-01-01

    The gummosis disease is caused by Botryosphaeria dothidea (Moug. ex. Fr) Ces. et de Not., and it is one of the most important diseases of stone fruits worldwide. The use of biocontrol as an alternative approach to synthetic chemical fungicides has aroused general concern about how to control plant diseases that are caused by phytopathogens. The aim of this study is to isolate Bacillus strains from raw honeys with the capacity to inhibit B. dothidea and to explore the mechanisms by which they could be used in the biocontrol of peach gummosis. Bacillus amyloliquefaciens SYBC H47 was isolated and identified on the basis of its physiological and biochemical characteristics and its 16S rRNA and gyrB gene sequences. The cell suspension and the cell-free supernatant of its culture showed significant antifungal activity against Aspergillus niger, Mucor racemosus, Fusarium oxysporum, Penicillium citrinum, and Candida albicans by agar-diffusion assays. The primary antifungal substances were bacillomycin L, fengycin, and surfactin, which were analyzed by HPLC LC/ESI-MS/MS. Bacillomycin L showed the best inhibitory effect against conidial germination of B. dothidea, followed by fengycin and surfactin. Surfactin had limited effects on mycelial growth, contrary to those of bacillomycin L and fengycin. However, a mixture of the three lipopeptides had a synergistic effect that disrupted the structure of the conidia and mycelia. In order to reduce the production cost, the use of waste frying peanut oil and soy oil as the sole carbon source increased the lipopeptide yield levels by approximately 17% (2.42 g/L) and 110% (4.35 g/L), respectively. In a field trial, the decreases in the infected gummosis rate (IGR) and the disease severity index (DSI) through cell suspension treatments were 20% and 57.5% (in 2014), respectively, and 40% and 57.5% (in 2015), respectively, in comparison with the control. In conclusion, B. amyloliquefaciens SYBC H47 could inhibit the germination of conidia

  18. A mutant of the nematophagous fungus Paecilomyces lilacinus (Thom) is a novel biocontrol agent for Sclerotinia sclerotiorum.

    Science.gov (United States)

    Yang, Fan; Abdelnabby, Hazem; Xiao, Yannong

    2015-12-01

    Sclerotinia sclerotiorum causes severe stem rot and yield loss in oilseed rape (Brassica napus L.) and other crops worldwide. Extensive studies have been conducted on Paecilomyces lilacinus as a nematophagous bioagent. However, no reports stated the effect of P. lilacinus as a biocontrol agent against oilseed rape rot S. sclerotiorum. This study describes such effect in lab and field trials using the new transformant pt361 derived from the wild strain P. lilacinus 36-1. Unlike the wild-type strain, the mutant pt361 showed high antagonistic effect against S. Sclerotiorum A. Under lab conditions, the pt361 inhibited (65%) radial mycelial growth of S. sclerotiorum in dual culture test producing 5.9 mm inhibition zone IZ in front of the S. sclerotiorum colony. Moreover, the cell-free filtrate of pt361 culture showed strong inhibitory effects (60.3-100%) on mycelial growth of S. sclerotiorum. In leaf detached assay, pt361 significantly (p rape stem rot, promoted growth and increased yield compared to the control and exceeded, at dose 100%, the action of the fungicide procymidone(®). In conclusion, the mutant pt361 of P. lilacinus is a novel and promising biocontrol agent against oilseed rape Sclerotinia stem rot. PMID:26521137

  19. Research Progress of Biocontrol Effect of Trichoderma harzianum%哈茨木霉生物防治研究进展

    Institute of Scientific and Technical Information of China (English)

    吕黎; 许丽媛; 罗志威; 周艳; 郭帅; 丰来

    2013-01-01

    木霉被认为是最有希望的生物防治因子,而哈茨木霉是木霉属中应用最为广泛的菌株。从哈茨木霉的生防效果、生防作用机制以及基因水平上的抗生机制等方面的研究进展进行了综述,并阐述了哈茨木霉可能的应用前景。%Trichoderma is the most promising isolated soil fungi for their ability to protect plants and contain pathogen populations under different soil conditions. While Trichoderma harzianum is the most commonly applied strain among Trichoderma. The biocontrol effect, the biocontrol mechanism, and the antibiotic mechanism at the gene level of Trichoderma harzianum were summarized, and the applying future of it was elaborated.

  20. Candida parapsilosis as a Potent Biocontrol Agent against Growth and Aflatoxin Production by Aspergillus Species

    Science.gov (United States)

    Niknejad, F; Zaini, F; Faramarzi, MA; Amini, M; Kordbacheh, P; Mahmoudi, M; Safara, M

    2012-01-01

    Background: Aflatoxin contamination of food and feed stuff is a serious health problem and significant economic concerns. In the present study, the inhibitory effect of Candida parapsilosis IP1698 on mycelial growth and aflatoxin production in aflatoxigenic strains of Aspergillus species was investigated. Methods: Mycelial growth inhibitions of nine strains of aflatoxigenic and non-aflatoxigenic Aspergillus species in the presence of C. parapsilosis investigated by pour plate technique at different pH, temperature and time of incubation. Reduction of aflatoxin was evaluated in co-cultured fungi in yeast extract sucrose broth after seven days of incubation using HPLC method. The data were analyzed by SPSS 11.5. Results: The presence of the C. parapsilosis at different pH did not affect significantly the growth rate of Aspergillus isolates. On the other hand, temperature and time of incubation showed to be significantly effective when compared to controls without C. parapsilosis (P≤0.05). In aflatoxigenic strains, minimum percentage of reductions in total aflatoxin and B1, B2, G1, G2 fractions were 92.98, 92.54, 77.48, 54.54 and 72.22 and maximum percentage of reductions were 99.59, not detectable, 94.42, and not detectable in both G1 and G2, respectively. Conclusion: C. parapsilosis might employ as a good biocontrol agent against growth and aflatoxin production by aflatoxigenic Aspergillus species PMID:23308351

  1. Candida Parapsilosis as a Potent Biocontrol Agent against Growth and Aflatoxin Production by Aspergillus Species

    Directory of Open Access Journals (Sweden)

    F Niknejad

    2012-10-01

    Full Text Available Background: Aflatoxin contamination of food and feed stuff is a serious health problem and significant economic concerns. In the present study, the inhibitory effect of Candida parapsilosis IP1698 on mycelial growth and aflatoxin production in aflatoxigenic strains of Aspergillus species was investigated.Methods: Mycelial growth inhibitions of nine strains of aflatoxigenic and non-aflatoxigenic Aspergillus species in thepresence of C. parapsilosis investigated by pour plate technique at different pH, temperature and time of incubation.Reduction of aflatoxin was evaluated in co-cultured fungi in yeast extract sucrose broth after seven days of incubation using HPLC method. The data were analyzed by SPSS 11.5.Results: The presence of the C. parapsilosis at different pH did not affect significantly the growth rate of Aspergillus isolates. On the other hand, temperature and time of incubation showed to be significantly effective when compared to controls without C. parapsilosis (P≤0.05. In aflatoxigenic strains, minimum percentage of reductions in total aflatoxin and B1, B2, G1, G2 fractions were 92.98, 92.54, 77.48, 54.54 and 72.22 and maximum percentage ofreductions were 99.59, not detectable, 94.42, and not detectable in both G1 and G2, respectively.Conclusion: C. parapsilosis might employ as a good biocontrol agent against growth and aflatoxin production by aflatoxigenic Aspergillus species.

  2. Phages of Listeria offer novel tools for diagnostics and biocontrol

    Directory of Open Access Journals (Sweden)

    Martin J Loessner

    2014-04-01

    Full Text Available Historically, bacteriophages infecting their hosts have perhaps been best known and even notorious for being a nuisance in dairy-fermentation processes. However, with the rapid progress in molecular microbiology and microbial ecology, a new dawn has risen for phages. This review will provide an overview on possible uses and applications of Listeria phages, including phage-typing, reporter phage for bacterial diagnostics, and use of phage as biocontrol agents for food safety. The use of phage-encoded enzymes such as endolysins for the detection and as antimicrobial will also be addressed. Desirable properties of candidate phages for biocontrol will be discussed. While emphasizing the enormous future potential for applications, we will also consider some of the intrinsic limitations dictated by both phage and bacterial ecology.

  3. Genome Sequencing and Comparative Analysis of the Biocontrol Agent Trichoderma harzianum sensu stricto TR274

    Energy Technology Data Exchange (ETDEWEB)

    Steindorff, Andrei S.; Noronha, Elilane F.; Ulhoa, Cirano J.; Kuo, Alan; Salamov, Asaf A.; Haridas, Sajeet; Riley, Robert W.; Druzhinina, Irina S.; Kubicek, Christian P.; Grigoriev, Igor V.

    2015-03-17

    Biological control is a complex process which requires many mechanisms and a high diversity of biochemical pathways. The species of Trichoderma harzianum are well known for their biocontrol activity against many plant pathogens. To gain new insights into the biocontrol mechanism used by T. harzianum, we sequenced the isolate TR274 genome using Illumina. The assembly was performed using AllPaths-LG with a maximum coverage of 100x. The assembly resulted in 2282 contigs with a N50 of 37033bp. The genome size generated was 40.8 Mb and the GC content was 47.7%, similar to other Trichoderma genomes. Using the JGI Annotation Pipeline we predicted 13,932 genes with a high transcriptome support. CEGMA tests suggested 100% genome completeness and 97.9% of RNA-SEQ reads were mapped to the genome. The phylogenetic comparison using orthologous proteins with all Trichoderma genomes sequenced at JGI, corroborates the Trichoderma (T. asperellum and T. atroviride), Longibrachiatum (T. reesei and T. longibrachiatum) and Pachibasium (T. harzianum and T. virens) section division described previously. The comparison between two Trichoderma harzianum species suggests a high genome similarity but some strain-specific expansions. Analyses of the secondary metabolites, CAZymes, transporters, proteases, transcription factors were performed. The Pachybasium section expanded virtually all categories analyzed compared with the other sections, specially Longibrachiatum section, that shows a clear contraction. These results suggests that these proteins families have an important role in their respective phenotypes. Future analysis will improve the understanding of this complex genus and give some insights about its lifestyle and the interactions with the environment.

  4. Environmental impact assessment of genetically modified biocontrol agents

    International Nuclear Information System (INIS)

    This review summarises the theoretical basis of risk analysis, and the political and social implications of introducing new biotechnology products in agricultural environments. The main factors to be considered under the present European regulation in the environmental impact assessment of genetically modified biocontrol agents are briefly discussed. Finally, an alternative risk assessment paradigm is proposed for genetically modified microorganisms, which shall consider the intrinsic properties of each antagonist, rather than the method used for generating it

  5. Biocontrol: Bacillus penetrans and Related Parasites of Nematodes

    OpenAIRE

    Sayre, R. M.

    1980-01-01

    Bacillus penetrans Mankau, 1975, previously described as Duboscqia penetrans Thorne 1940, is a candidate agent for biocontrol of nematodes. This review considers the life stages of this bacterium: vegetative growth phase, colony fragmentation, sporogenesis, soil phase, spore attachment, and penetration into larvae of root-knot nematodes. The morphology of the microthallus colonies and the unusual external features of the spore are discussed. Taxonomic affinities with the actinomycetes, partic...

  6. Environmental and molecular regulation of phenazine-1-carboxamide biosynthesis in Pseudomonas chlororaphis strain PCL1391

    NARCIS (Netherlands)

    Rij, Evert Tjeerd van

    2006-01-01

    The biocontrol strain studied in this thesis is Pseudomonas chlororaphis strain PCL1391, which controls tomato foot and root rot caused by Fusarium oxysporum f. sp. radicis-licopersici (Chin-A-Woeng et al., 1998). Production of the anti-fungal metabolite phenazine-1-carboxamide (PCN) and an efficien

  7. Cucumber Rhizosphere Microbial Community Response to Biocontrol Agent Bacillus subtilis B068150

    Directory of Open Access Journals (Sweden)

    Lihua Li

    2015-12-01

    Full Text Available Gram-positive bacteria Bacillus subtilis B068150 has been used as a biocontrol agent against the pathogen Fusarium oxysporum cucumerinum. Cucumber was grown in three soils with strain B068150 inoculated in a greenhouse for 90 days, and the colonization ability of strain B068150 in cucumber rhizosphere and non-rhizosphere soils was determined. Changes in total bacteria and fungi community composition and structures using denaturing gradient gel electrophoresis (DGGE and sequencing were determined. Colony counts showed that B068150 colonization in the rhizosphere was significantly higher (p < 0.001 than in non-rhizosphere soils. Based on our data, the introduction of B. bacillus B068150 did not change the diversity of microbial communities significantly in the rhizosphere of three soils. Our data showed that population density of B068150 in clay soil had a significant negative correlation on bacterial diversity in cucumber rhizosphere in comparison to loam and sandy soils, suggesting that the impact of B068150 might be soil specific.

  8. Improved delivery of biocontrol Pseudomonas and their antifungal metabolites using alginate polymers.

    Science.gov (United States)

    Russo, A; Moënne-Loccoz, Y; Fedi, S; Higgins, P; Fenton, A; Dowling, D N; O'Regan, M; O'Gara, F

    1996-02-01

    Alginate polymer was evaluated as a carrier for seed inoculation with a genetically modified strain Pseudomonas fluorescens F113LacZY, which protects sugar-beet against Pythium-mediated damping-off. F113LacZY survived in alginate beads at 5 log10 CFU/bead or higher counts for 8 weeks of storage, regardless of the conditions of incubation. In plant inoculation experiments, colonisation of the growing area of the root by F113LacZY, derived from alginate beads placed in the soil next to the seed or from an alginate coating around the seeds, was improved compared with application of just free cells of the strain. F113LacZY trapped in alginate beads was an effective producer of antifungal phloroglucinols as indicated by direct HPLC quantification of phloroglucinols and in vitro inhibition of both the indicator bacterium Bacillus subtilis A1 and the pathogenic fungus Pythium ultimum. Alginate polymer represents a promising carrier for the delivery of biocontrol inoculants for root colonisation and production of antifungal metabolites. PMID:8867631

  9. The relationship of biofilm production to biocontrol activity of Burkholderia pyrrocinia FP62

    Science.gov (United States)

    Foliar biocontrol agent (BCA) efficacy is often inconsistent due to poor colonization and survival on plant surfaces. Burkholderia pyrrocinia FP62, a superior leaf colonist and BCA of Botrytis cinerea, forms unsaturated biofilms on plant surfaces. To determine the relationship between biocontrol act...

  10. Advances in biocontrol mechanism and application of Trichoderma spp. for plant diseases

    Institute of Scientific and Technical Information of China (English)

    HUANG Caihong; YANG Qian

    2007-01-01

    Trichoderma spp. is a filamentous soil fungus known as an effective biocontrol agent of a range of important airborne and soilborne pathogens, it has universal distribution and economic importance. This article reviewed the researches on biocontrol mechanism for plant diseases and application of Trichoderma spp., especially Trichoderma harzianum in recent years.

  11. Yeast microflora of nectarines and their potential for biocontrol of brown rot

    Science.gov (United States)

    Resident fruit microflora has been the source of biocontrol agents for the control of postharvest decays of fruits and the active ingredient in commercialized biocontrol products. With the exception of grapes and apples, information on the resident microflora of other fruits is only fragmentary; ho...

  12. Efficacy of Pseudomonas fluorescens for biocontrol of Escherichia coli 0157:H7 on spinach

    Science.gov (United States)

    Control of foodborne pathogens on leafy green vegetables is crucial for consumer food safety. Biocontrol microbes may inhibit or suppress foodborne pathogen growth and enhance existing post-harvest intervention controls. The efficacy of Pseudomonas fluorescens for biocontrol of Escherichia coli O157...

  13. Genetic responses induced in olive roots upon colonization by the biocontrol endophytic bacterium Pseudomonas fluorescens PICF7.

    Directory of Open Access Journals (Sweden)

    Elisabetta Schilirò

    Full Text Available Knowledge on the genetic basis underlying interactions between beneficial bacteria and woody plants is still very limited, and totally absent in the case of olive. We aimed to elucidate genetic responses taking place during the colonization of olive roots by the native endophyte Pseudomonas fluorescens PICF7, an effective biocontrol agent against Verticillium wilt of olive. Roots of olive plants grown under non-gnotobiotic conditions were collected at different time points after PICF7 inoculation. A Suppression Subtractive Hybridization cDNA library enriched in induced genes was generated. Quantitative real time PCR (qRT-PCR analysis validated the induction of selected olive genes. Computational analysis of 445 olive ESTs showed that plant defence and response to different stresses represented nearly 45% of genes induced in PICF7-colonized olive roots. Moreover, quantitative real-time PCR (qRT-PCR analysis confirmed induction of lipoxygenase, phenylpropanoid, terpenoids and plant hormones biosynthesis transcripts. Different classes of transcription factors (i.e., bHLH, WRKYs, GRAS1 were also induced. This work highlights for the first time the ability of an endophytic Pseudomonas spp. strain to mount a wide array of defence responses in an economically-relevant woody crop such as olive, helping to explain its biocontrol activity.

  14. Screening of bacterial biocontrols against sapstain fungus (Lasiodiplodia theobromae Pat.) of rubberwood (Hevea brasiliensis Muell.Arg.).

    Science.gov (United States)

    Sajitha, K L; Maria Florence, E J; Dev, Suma Arun

    2014-09-01

    Diverse bacterial biocontrol agents from various sources of aerobic composts against the sapstain fungus Lasiodiplodia theobromae in rubberwood (Hevea brasiliensis) were isolated, screened and identified by various morphological, biochemical and molecular techniques. The inhibitory effect of seventeen bacterial isolates was examined and seven exhibited inhibition towards the sapstain fungus. Among the seven antagonists, six were conclusively identified as Bacillus subtilis and one as Paenibacillus polymyxa using 16S rRNA-encoding gene sequencing. This is the first report on the occurrence of P. polymyxa, a potent biofertilizer and antagonist in vermicompost. HiCrome Bacillus agar was identified as an effective medium for differentiation of B. subtilis from other Bacillus species. The present work demonstrates the efficacy of the antagonistic property of B. subtilis strains against rubberwood sapstain fungus. Culture-based antagonistic inhibition displayed by B. subtilis can be extended to cater to the biocontrol requirements of wood-based industries against the stain fungus. The study showed the utility of an integrated approach, employing morphological, biochemical and molecular tools for conclusive identification of several bacterial isolates present in aerobic composts from diverse sources. PMID:25049165

  15. Potencialidade antagonística detectada em alguns procariotas agentes de biocontrole de enfermidades de plantas Antagonistic potentiality in prokaryotic biocontrol agents for plant diseases

    Directory of Open Access Journals (Sweden)

    Victor Rafael Barra

    2008-06-01

    Full Text Available Ao longo de vários anos, no Laboratório de Bacteriologia de Plantas e Controle Biológico (LBPCB do Departamento de Fitopatologia da Universidade Federal de Viçosa têm-se, sistematicamente, isolado, de rizosfera, rizoplano e filoplano de plantas cultivadas de importância econômica, milhares de procariotas os quais são testados, um a um, como agentes de biocontrole de enfermidades. A grande maioria deles, como esperado, não exibe qualquer potencialidade como agente de biocontrole e é descartada. Alguns poucos são selecionados, através de demorados ensaios de biocontrole experimental tanto em casa de vegetação como em campo. Neste trabalho, mostra-se que muitos dos organismos em estudo, mesmo havendo sido selecionados, principalmente, por induzirem resistência sistêmica na planta alvo, expressam constitutivamente mecanismos de antagonismo microbiano que parecem fazer parte de seu perfil enquanto agentes de biocontrole.Along several years, at the Laboratory of Plant Bacteriology and Biological Control of the Department of Plant Pathology of the Federal University of Viçosa, thousands of prokaryote isolates have been obtained from the rhizosphere, rhizoplane and phylloplane of economically important plants. Every obtained isolate is tested as biocontrol agents. As expected, the majority does not show any biocontrol potentiality and is discarded. Few are selected through time-consuming experimental biocontrol trials in greenhouse and field. Even being selected mainly for their ability to induce resistance in target plants, many of them constitutively express mechanisms of antagonism against pathogens that seem to be part of their performance while biocontrol agents.

  16. The Potential Use of Wolbachia-Based Mosquito Biocontrol Strategies for Japanese Encephalitis.

    Directory of Open Access Journals (Sweden)

    Claire L Jeffries

    Full Text Available Japanese encephalitis virus (JEV is a zoonotic pathogen transmitted by the infectious bite of Culex mosquitoes. The virus causes the development of the disease Japanese encephalitis (JE in a small proportion of those infected, predominantly affecting children in eastern and southern Asia. Annual JE incidence estimates range from 50,000-175,000, with 25%-30% of cases resulting in mortality. It is estimated that 3 billion people live in countries in which JEV is endemic. The virus exists in an enzootic transmission cycle, with mosquitoes transmitting JEV between birds as reservoir hosts and pigs as amplifying hosts. Zoonotic infection occurs as a result of spillover events from the main transmission cycle. The reservoir avian hosts include cattle egrets, pond herons, and other species of water birds belonging to the family Ardeidae. Irrigated rice fields provide an ideal breeding ground for mosquitoes and attract migratory birds, maintaining the transmission of JEV. Although multiple vaccines have been developed for JEV, they are expensive and require multiple doses to maintain efficacy and immunity. As humans are a "dead-end" host for the virus, vaccination of the human population is unlikely to result in eradication. Therefore, vector control of the principal mosquito vector, Culex tritaeniorhynchus, represents a more promising strategy for reducing transmission. Current vector control strategies include intermittent irrigation of rice fields and space spraying of insecticides during outbreaks. However, Cx. Tritaeniorhynchus is subject to heavy exposure to pesticides in rice fields, and as a result, insecticide resistance has developed. In recent years, significant advancements have been made in the potential use of the bacterial endosymbiont Wolbachia for mosquito biocontrol. The successful transinfection of Wolbachia strains from Drosophila flies to Aedes (Stegomyia mosquitoes has resulted in the generation of "dengue-refractory" mosquito

  17. Isolation and characterization of antagonistic Bacillus strains capable to degrade ethylenethiourea.

    Science.gov (United States)

    Vágvölgyi, Csaba; Sajben-Nagy, Enikő; Bóka, Bettina; Vörös, Mónika; Berki, Adrienn; Palágyi, Andrea; Krisch, Judit; Skrbić, Biljana; Durišić-Mladenović, N; Manczinger, László

    2013-03-01

    In this study, more than 150 bacteria showing antagonistic properties against bacterial and fungal pathogens of the tomato plant were isolated and characterized. The most efficient agents against these phytopathogenic microorganisms belong to the genus Bacillus: the best biocontrol isolates were representatives of Bacillus subtilis, B. mojavensis and B. amyloliquefaciens species. They intensively produced fengycin or/and surfactin depsipeptide antibiotics and also proved to be excellent protease secretors. It was proved, that the selected strains were able to use ethylenethiourea (ETU) as sole nitrogen source. These antagonistic and ETU-degrading Bacillus strains can be applied as biocontrol and also as bioremediation agents. PMID:23143288

  18. 用于土壤中生防芽孢杆菌B006和BH1检测的SCAR标记及其特异性%Identification of the SCAR Markers for Detection of Biocontrol Bacillus Strains B006 and BH1 in Soil

    Institute of Scientific and Technical Information of China (English)

    耿文义; 郭荣君; 李世东; 许修宏

    2011-01-01

    Two Bacillus strains, B006 and BH1, are highly suppressive against Fusarium spp., the causal agents of plant root diseases. Discrimination of these strains from native soil bacteria was critical to study their population dynamics and behaviors in soil. Polymorphism in genomic DNA of the strains B006 and BH1 as well as 25 references including Bacillus spp., Sphingosine spp. And Pseudomonas spp. Were analyzed with 16 random primer pairs by RAPD. Sequence-specific fragments with 730 bp and 658 bp could be obtained from the strains B006 and BH1 by PCR with single specific primers B19 and C01, respectively, which were absent in other strains, hi order to increase the amplification specificity and stability, 6 and 12 SCAR marker primer pairs were designed on the base of the nucleotide sequence information of RAPDs closely linked to the strains B006 and BH1, respectively, and their specificity were confirmed by amplification of specific DNA bands of the strains B006 and BH1. Two primer sets SCAR-B006 and SCAR-BH1 were selected and the amplification products with 523 bp and 658 bp were transformed into more-specific SCAR markers, respectively. Specificity of these markers was further tested in natural soil. Only the soil amended with B006 or BH1 gave specific SCAR bands. This PCR-based method is useful to track the strains B006 and BH1 in soil samples rapidly and efficiently.%生防芽孢杆菌Bacillus spp.菌株B006和BH1对引发多种根腐病的真菌具有拮抗作用,研发特异性标记是研究该菌株在土壤中生态行为的关键.本研究采用16个RAPD随机引物对B006和BH1以及包括芽孢杆菌、鞘氨醇杆菌Sphingosinc spp.和假单胞菌Pseudomonas spp.在内的25株参试菌株的基囚组DNA多态性进行了分析,其中利用单引物B19和C01可分别扩增到仅出现于B006和BH1基因图谱中的特异性条带(大小分别为730bp和658bp).为提高扩增的特异性和稳定性,根据B006和BH1的RAPD标记中

  19. Introducing the term 'Biocontrol Plants' for integrated pest management

    Directory of Open Access Journals (Sweden)

    Pia Parolin

    2014-02-01

    Full Text Available Studies of interactions between crops, additional plants, pests and beneficial organisms already exist as well as studies of natural enemy preference, dispersal, and abundance. However, these studies focus on tri-trophic interactions from an "arthropod" point of view. We think that in order to optimize crop protection methods we need to understand the effects that plant structures have on the various arthropods and on subsequent tri-trophic interactions. Although studies and reviews describing the role of secondary plants in Integrated Pest Management (IPM exist, to date a general term which encompasses all plants added to a cropping system with the aim of enhancing IPM strategies has yet to be formulated. Therefore, we suggest a new term, "biocontrol plants", which we define as plants that are intentionally added to a crop system with the aim of enhancing crop productivity through pest attraction and/or pest regulation; a term that will promote the use of biocontrol services, and can ultimately lead to an increase in the sustainability of cropping systems.

  20. Selection and Characterization of Endophytic Bacteria as Biocontrol Agents of Tomato Bacterial Wilt Disease

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    ABDJAD ASIH NAWANGSIH

    2011-06-01

    Full Text Available Biological control of bacterial wilt pathogen (Ralstonia solanacearum of tomato using endophytic bacteria is one of the alternative control methods to support sustainable agriculture. This study was conducted to select and characterize endophytic bacteria isolated from healthy tomato stems and to test their ability to promote plant growth and suppress bacterial wilt disease. Among 49 isolates successfully isolated, 41 were non-plant pathogenic. Green house test on six selected isolates based on antagonistic effect on R. solanacearum or ability to suppress R. solanacearum population in dual culture assays obtained BC4 and BL10 isolates as promising biocontrol agents. At six weeks after transplanting, plants treated with BC4 isolate showed significantly lower disease incidence (33% than that of control (83%. Plants height was not significantly affected by endophytic bacterial treatments. Based on 16S rRNA sequence, BC4 isolate had 97% similarity with Staphylococcus epidermidis (accession number EU834240.1, while isolate BL10 had 98% similarity with Bacillus amyloliquefaciens strain JK-SD002 (accession number AB547229.1.

  1. The biocontrol endophytic bacterium Pseudomonas fluorescens PICF7 induces systemic defense responses in aerial tissues upon colonization of olive roots

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    Carmen eGómez-Lama Cabanás

    2014-09-01

    Full Text Available Pseudomonas fluorescens PICF7, a native olive root endophyte and effective biocontrol agent (BCA against Verticillium wilt of olive, is able to trigger a broad range of defense responses in root tissues of this woody plant. In order to elucidate whether strain PICF7 also induces systemic defense responses in above-ground organs, aerial tissues of olive plants grown under non-gnotobiotic conditions were collected at different time points after root bacterization with this endophytic BCA. A suppression subtractive hybridization (SSH cDNA library, enriched in up-regulated genes, was generated. This strategy enabled the identification of 376 ESTs (99 contigs and 277 singlets, many of them related to response to different stresses. Five ESTs, involved in defense responses, were selected to carry out time-course quantitative real-time PCR (qRT-PCR experiments aiming to: (i validate the induction of these genes, and (ii shed light on their expression pattern along time (from 1 to 15 days. Induction of olive genes potentially coding for lypoxigenase 2, catalase, 1-aminocyclopropane-1-carboxylate oxidase and phenylananine ammonia-lyase was thus confirmed at some time points. Computational analysis also revealed that different transcription factors were up-regulated in olive aerial tissues (i.e. jerf, bHLH, WRKYs, as previously reported for roots. Results confirmed that root colonization by this endophytic bacterium does not only trigger defense responses in this organ but also mount a wide array of systemic defense responses in distant tissues (stems, leaves. This sheds light on how olive plants respond to the ‘non-hostile’ colonization by a bacterial endophyte and how induced defense response can contribute to the biocontrol activity of strain PICF7.

  2. Bacillus amyloliquefaciens GA1 as a source of potent antibiotics and other secondary metabolites for biocontrol of plant pathogens

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    Brans Alain

    2009-11-01

    Full Text Available Abstract Background Phytopathogenic fungi affecting crop and post-harvested vegetables are a major threat to food production and food storage. To face these drawbacks, producers have become increasingly dependent on agrochemicals. However, intensive use of these compounds has led to the emergence of pathogen resistance and severe negative environmental impacts. There are also a number of plant diseases for which chemical solutions are ineffective or non-existent as well as an increasing demand by consumers for pesticide-free food. Thus, biological control through the use of natural antagonistic microorganisms has emerged as a promising alternative to chemical pesticides for more rational and safe crop management. Results The genome of the plant-associated B. amyloliquefaciens GA1 was sample sequenced. Several gene clusters involved in the synthesis of biocontrol agents were detected. Four gene clusters were shown to direct the synthesis of the cyclic lipopeptides surfactin, iturin A and fengycin as well as the iron-siderophore bacillibactin. Beside these non-ribosomaly synthetised peptides, three additional gene clusters directing the synthesis of the antibacterial polyketides macrolactin, bacillaene and difficidin were identified. Mass spectrometry analysis of culture supernatants led to the identification of these secondary metabolites, hence demonstrating that the corresponding biosynthetic gene clusters are functional in strain GA1. In addition, genes encoding enzymes involved in synthesis and export of the dipeptide antibiotic bacilysin were highlighted. However, only its chlorinated derivative, chlorotetaine, could be detected in culture supernatants. On the contrary, genes involved in ribosome-dependent synthesis of bacteriocin and other antibiotic peptides were not detected as compared to the reference strain B. amyloliquefaciens FZB42. Conclusion The production of all of these antibiotic compounds highlights B. amyloliquefaciens GA1 as

  3. Improving biocontrol activity of Pseudomonas fluorescens through chromosomal integration of 2,4-diacetylphloro- glucinol biosynthesis genes

    Institute of Scientific and Technical Information of China (English)

    ZHOU Hongyou; WEI Hailei; LIU Xili; WANG Ye; ZHANG Liqun; TANG Wenhua

    2005-01-01

    Antibiotic 2,4-diacetylphloroglucinol (2,4- DAPG) produced by Pseudomonas fluorescens CPF-10 and 2P24 is a principal factor enabling bacteria to suppress plant diseases caused by soilborne pathogens. In this study, a 2,4-DAPG biosynthesis locus phlACBDE cloned from strain CPF-10 was assembled into a mini-Tn5 transposon and introduced into the chromosome of P. fluorescens P32 (2,4- DAPG-), CPF-10 and 2P24 to construct the 2,4-DAPG overproducing derivatives P32-38, CPF10-9 and 2P24-48, respectively. All the transgenic strains showed an enhanced antibiosis capacity against plant microbial pathogens in vitro and two strains, P32-38 and CPF10-9, provided significantly better protection against wheat take-all disease caused by Gaeumannomyces graminis var. tritici and tomato bacterial wilt caused by Ralstonia solanacearum in greenhouse. Compared to their parental strains, the 2,4-DAPG overproducing derivatives colonized to the same extent on the wheat tips in the autoclaved soil, but developed larger populations in natural soil. These results indicated that production of antibiotics 2,4- DAPG by biological control pseudomonads can contribute not only to their disease suppression capacities but also to the ecological competence in the resident microflora. Our research also suggests that it is a realistic approach to improve biocontrol capacity of P. fluorescens through the genetic modification of its antibiotic 2,4-DAPG production.

  4. Risk assessment: progress of quarantine biocontrol research on Chinese Tallow, Melaleuca, and Downy Rose Myrtle

    Science.gov (United States)

    Risk assessments of two biocontrol candidates for Chinese tallow, Triadica sebifera (Euphoriales: Euphorbiaceae), and one for Melaleuca, Melaleuca quinquenervia (Myrtales: Myrtaceae), were conducted during 2009 and continuing into 2010 by USDA scientists located at the Florida Department of Agricul...

  5. Differential effects of organic compounds on cucumber damping-off and biocontrol activity of antagonistic bacteria

    DEFF Research Database (Denmark)

    Li, Bin; Ravnskov, Sabine; Guanlin, X.;

    2011-01-01

    The influence of the organic compounds tryptic soy broth, cellulose, glucose and chitosan on cucumber damping-off caused by Pythium aphanidermatum and biocontrol efficacy of the biocontrol agents (BCAs) Paenibacillus macerans and P. polymyxa were examined in a seedling emergence bioassay. Results...... showed that the organic compounds differentially affected both pathogen and BCAs. Tryptic soy broth, glucose and chitosan increased Pythium damping-off of cucumber, compared to the control treatment without organic compounds, whereas cellulose had no effect. Both Paenibacillus species had biocontrol...... effects against Pythium damping-off compared with the corresponding treatments with P. aphanidermatum alone, but the biocontrol efficacy depended on the type of organic compounds added. Both BCAs counteracted damping-off in treatments with TSB and chitosan. However, P. polymyxa counteracted damping...

  6. Detection and quantification by PCR assay of the biocontrol agent Pantoea agglomerans CPA-2 on apples.

    Science.gov (United States)

    Soto-Muñoz, Lourdes; Teixidó, Neus; Usall, Josep; Viñas, Inmaculada; Torres, Rosario

    2014-04-01

    The registration of biological control agents requires the development of monitoring systems to detect and quantify the agent in the environment. Pantoea agglomerans CPA-2 is an effective biocontrol agent for postharvest diseases of citrus and pome fruits. The monitoring of CPA-2 in postharvest semi-commercial trials was evaluated by Rodac impression plates and the colonies isolated were confirmed by conventional PCR using the SCAR primers PAGA1 and PAGB1. Samples were taken from different surfaces that had contact with CPA-2, the surrounding environment and working clothes worn by handlers. Moreover, population dynamics of the strain CPA-2 were determined on apple surfaces using both the classical plating technique and real-time quantitative PCR (qPCR). A qPCR assay using a 3'-minor groove-binding (MGB) probe was developed for the specific detection and quantification of P. agglomerans strain CPA-2. Based on the nucleotide sequence of a SCAR fragment of CPA-2, one primer set and TaqMan MGB probe were designed. The primers SP2-F/SP2-R and the TaqMan MGB probe showed a specific detection of strain CPA-2 on apple surfaces, which was verified tested against purified DNA from 17 strains of P. agglomerans, 4 related Pantoea species, and 21 bacterial strains from other genera isolated from whole and also freshly-cut fruit and vegetables. The detection level was approximately 10(3) cells per reaction, and the standard curve was linear within a range of 5log units. Results from semi-commercial trials showed that CPA-2 had a low impact. The maximum persistence of P. agglomerans CPA-2 was not longer than 5days in plastic boxes stored at 0°C. Significant differences in CPA-2 population level dynamics were observed in results obtained by qPCR and dilution plating. These differences may indicate the presence of non-degraded DNA from non-viable cells. In conclusion, qPCR is a novel potential tool to quickly and specifically monitor recent surface colonisation by CPA-2

  7. Endophytic Bacteria as Biocontrol Agents of Tomato Bacterial Wilt Disease

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    Arika Purnawati

    2014-01-01

    Full Text Available Ralstonia solanacearum is a tomato pathogen causing wilt disease which can lose crop production up to 100%. This problem also has economical impact to Indonesia, though controlling this pathogen has been donebefore. Biological control using endophytic bacteria is one of alternative control methods to support agriculture sustainability. The objective of these experiments are isolating endophytic bacteria taken from healthy tomato stems and root as well as investigating patogens as biocontrol agents of tomato bacterial wilt disease. Among 10 isolates found, only Ps1 and Ps8 can inhibit R. solanacearum based on antagonistic test in vitro using seed coat method and 4-7 mm inhibition zone forming. In vivo test was applied in green house in 30 days old of tomatoes usingendophytic bacteria by theroot soak technique. The result shows Ps1 and Ps8 within 15-16 days incubation period have suppressed 8.07%-9.19% pathogen attack.

  8. Chitinases from the Plant Disease Biocontrol Agent, Stenotrophomonas maltophilia C3.

    Science.gov (United States)

    Zhang, Z; Yuen, G Y; Sarath, G; Penheiter, A R

    2001-02-01

    ABSTRACT Stenotrophomonas maltophilia strain C3, a biocontrol agent of Bipolaris sorokiniana in turfgrass, produced chitinases in broth media containing chitin. Chitinases were partially purified from culture fluid by ammonium sulfate precipitation and chitin affinity chromatography. The chromatography fraction with the highest specific chitinase activity was inhibitory to conidial germination and germ-tube elongation of B. sorokiniana, but it was less inhibitory than the protein fraction or the raw culture filtrate. The fraction exhibited strong exochitinase and weak endo-chitinase activity. Optimum temperature and pH for chitinase activity were 45 to 50 degrees C and 4.5 to 5.0, respectively. Chitinase activity was inhibited by Hg(2+) and Fe(3+), but not by other metal ions or enzyme inhibitors. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the chromatography fraction revealed the presence of five protein bands of 25, 32, 48, 65, and 75 kDa. Partial amino acid sequences of the 32-, 65-, and 75-kDa proteins indicated that they are homologous to known bacterial chitinases. There was no homology found in the partial amino acid sequences of the 25- and 48-kDa proteins to any known chitinases. Five chitinase-active proteins were detected in the protein and chromatography fractions by activity gels, but when each protein was extracted and re-electrophoresed separately under denaturing conditions, only 32- or 48-kDa proteins were revealed. It was concluded that strain C3 produces at least two chitinases that are antifungal.

  9. Biocontrol of larval mosquitoes by Acilius sulcatus (Coleoptera: Dytiscidae

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    Banerjee Siddhartha S

    2008-10-01

    Full Text Available Abstract Background Problems associated with resistant mosquitoes and the effects on non-target species by chemicals, evoke a reason to find alternative methods to control mosquitoes, like the use of natural predators. In this regard, aquatic coleopterans have been explored less compared to other insect predators. In the present study, an evaluation of the role of the larvae of Acilius sulcatus Linnaeus 1758 (Coleoptera: Dytiscidae as predator of mosquito immatures was made in the laboratory. Its efficacy under field condition was also determined to emphasize its potential as bio-control agent of mosquitoes. Methods In the laboratory, the predation potential of the larvae of A. sulcatus was assessed using the larvae of Culex quinquefasciatus Say 1823 (Diptera: Culicidae as prey at varying predator and prey densities and available space. Under field conditions, the effectiveness of the larvae of A. sulcatus was evaluated through augmentative release in ten cemented tanks hosting immatures of different mosquito species at varying density. The dip density changes in the mosquito immatures were used as indicator for the effectiveness of A. sulcatus larvae. Results A single larva of A. sulcatus consumed on an average 34 IV instar larvae of Cx. quinquefasciatus in a 24 h period. It was observed that feeding rate of A. sulcatus did not differ between the light-on (6 a.m. – 6 p.m., and dark (6 p.m. – 6 a.m. phases, but decreased with the volume of water i.e., space availability. The prey consumption of the larvae of A. sulcatus differed significantly (P A. sulcatus larvae, while with the withdrawal, a significant increase (p A. sulcatus in regulating mosquito immatures. In the control tanks, mean larval density did not differ (p > 0.05 throughout the study period. Conclusion the larvae of the dytiscid beetle A. sulcatus proved to be an efficient predator of mosquito immatures and may be useful in biocontrol of medically important mosquitoes.

  10. Biocontrol of fouling pests: Effect of diversity, identity and density of control agents.

    Science.gov (United States)

    Atalah, Javier; Newcombe, Emma M; Zaiko, Anastasija

    2016-04-01

    Augmentative biocontrol, using native natural enemies, has been suggested as a promising tool to control marine biofouling pests on artificial structures. However, there are still important knowledge gaps to be addressed before biocontrol can be considered as a management tool. In a field experiment on floating marine structures we examined intra- and interspecific consumer interactions among biocontrol agents on different surface orientations. We tested the effect of identity, density and diversity of three invertebrates (the 11-arm seastar Coscinasterias muricata, the sea urchin Evechinus chloroticus and the gastropod Cook's turban Cookia sulcata) to reduce established biofouling and to prevent fouling growth on defouled surfaces. High densities of biocontrol agents were not more effective at fouling control (cover and biomass) than low densities. Nor did multi-species treatments function more effectively than mono-specific ones. However, biocontrol agent identity was important, with the 11-arm seastar and Cook's turban being the most effective at fouling reduction and prevention, respectively. Surface orientation had a strong effect on the effectiveness of control agents, with the best results obtained on vertical compared to diagonal and underside surfaces. This study confirmed the potential of biocontrol as a management tool for marine pest, indicating that identity is more important than richness and density of control agents. It also highlighted the limitations of this approach on diagonal and underside surfaces, where control agents have limited retention ability. PMID:26845376

  11. 三株生防酵母菌对Botrytis cinerea的抑菌作用研究%Study on antifungal activities of biocontrol yeasts against Botrytis cinerea

    Institute of Scientific and Technical Information of China (English)

    王傲雪; 关鑫; 张俊峰; 张珍珠; 王瑞虎; 陈秀玲

    2014-01-01

    In this study, the antifungal activities of biocontrol yeasts Cryptococcus albidus 63(Ca63), Cryptococcus albidus 64 (Ca64) and Candida parapsilosis yett1006 against Botrytis cinerea were investigated. The dual culture between three biocontrol yeasts and B. cinerea t08016b results showed that no obvious inhibiting zone appeared, but the pathogen growed more weakly than the control. The results of chitinase andβ-1, 3-glucanase activities assay showed that the levels of the two enzymes activities did not change significantly (P>0.05) when the three biocontrol yeasts cultured with pathogen. The inhibition rates of three biocontrol yeast strains on mycelium growth were 88.57%, 90.66% and 85.65%, respectively. High concentrations of biocontrol yeasts had a strong inhibitory effect on the spore germination of B. cinerea t08016b. Especial y, 1 × 108 cfu·mL-1 biocontrol yeasts almost stopped the spore germination of B. cinerea t08016b. Heat-inactived yeast suspension and culture supernatant had no significant inhibitory effects on B. cinerea. These results il ustrated that the antagonic effect of biocontrol yeasts was mainly due to the competition of limited space and nutrient conditions, not secreting antifungal substances.%针对三株生防酵母菌Cryptococcus albidus 63(Ca63)、Cryptococcus albidus 64(Ca64)、Candida parapsilosis yett1006对番茄灰霉病病原菌B. cinerea t08016b的抑菌作用进行研究。三株生防酵母菌与B. cinerea t08016b的对峙培养过程中,未产生明显抑菌圈,均可减弱B. cinerea t08016b生长势;几丁质酶和β-1,3-葡聚糖酶活性测定结果表明, t08016b并无诱导三株酵母菌分泌酶活显著增加(P>0.05);研究酵母菌对B. cinerea t08016b菌丝生长和孢子萌发率影响发现,C.albidus 63、C.albidus 64和C.parapsilosis yett1006对菌丝生长量的抑制率分别为88.57%、90.66%和85.65%,高浓度菌液对孢子萌发有显著抑制效果,1×108 cfu·mL-1菌体活

  12. Mutation of rpiA in Enterobacter cloacae decreases seed and root colonization and biocontrol of damping-off caused by Pythium ultimum on cucumber.

    Science.gov (United States)

    Lohrke, Scott M; Dery, Pierre D; Li, Wei; Reedy, Ralph; Kobayashi, Donald Y; Roberts, Daniel R

    2002-08-01

    Strains of Enterobacter cloacae show promise as biocontrol agents for Pythium ultimum-induced damping-off on cucumber and other crops. E. cloacae A145 is a mini-Tn5 Km transposon mutant of strain 501R3 that was significantly reduced in suppression of damping-off on cucumber caused by P. ultimum. Strain A145 was deficient in colonization of cucumber, sunflower, and wheat seeds and significantly reduced in colonization of corn and cowpea seeds relative to strain 501R3. Populations of strain A145 were also significantly lower than those of strain 501R3 at all sampling times in cucumber, wheat, and sunflower rhizosphere. Populations of strain A145 were not detectable in any rhizosphere after 42 days, while populations of strain 501R3 remained at substantial levels throughout all experiments. Molecular characterization of strain A145 indicated mini-Tn5 Km was inserted in a region of the E. cloacae genome with a high degree of DNA and amino acid sequence similarity to rpiA, which encodes ribose-5-phosphate isomerase. In Escherichia coli, RpiA catalyzes the interconversion of ribose-5-phosphate and ribulose-5-phosphate and is a key enzyme in the pentose phosphate pathway. Ribose-5-phosphate isomerase activity in cell lysates from strain A145 was approximately 3.5% of that from strain 501R3. In addition, strain A145 was a ribose auxotroph, as expected for an rpiA mutant. Introduction of a 1.0-kb DNA fragment containing only the rpiA homologue into strain A145 restored ribose phosphate isomerase activity, prototrophy, seedling colonization, and disease suppression to levels similar to those associated with strain 501R3. Experiments reported here indicate a key role for rpiA and possibly the pentose phosphate pathway in suppression of damping-off and colonization of subterranean portions of plants by E. cloacae. PMID:12182339

  13. Biocontrole de Listeria monocytogenes por Pediococcus acidilactici em couve minimamente processada Biocontrol of Listeria monocytogenes by Pediococcus acidilactici in fresh-cut kale

    Directory of Open Access Journals (Sweden)

    Wanessa Altimiras Costa

    2009-12-01

    Full Text Available Este estudo avaliou um sistema de biocontrole para inibição de Listeria monocytogenes em couve minimamente processada, objetivando sua segurança durante estocagem sob refrigeração e em condições de abuso de temperatura. O potencial inibitório de bactérias láticas tolerantes ao sal e psicrotróficas contaminantes naturais da couve e Lactobacillus plantarum, Lactobacillus delbrueckii ATCC 9649 e Lactobacillus casei CCT 1465 foram avaliadas contra L. monocytogenes. O isolado de couve identificado como P. acidilactici CCA3 inibiu L. monocytogenes a 10 e 15 °C em ágar MRS e foi selecionado como possível agente de biocontrole. O número de L. monocytogenes na couve minimamente processada aumentou 3,7 e 4,7 ciclos logarítmicos a 5 e 10 °C, respectivamente, após 20 dias de armazenamento e 4,6 ciclos logarítmicos após oito dias a 15 °C. Entretanto, quando 10(8 UFC.g-1 de P. acidilactici CCA3 foram inoculados no produto processado, o crescimento de L. monocytogenes reduziu 2,3 ciclos logarítmicos sob temperatura abusiva de 15 °C. A acidez titulável e as características sensoriais da couve não foram alteradas pela presença de CCA3 ao longo do período de vida útil. Estes resultados sugerem o potencial de aplicação dos bioconservantes na couve minimamente processada, que necessitam estar associados à refrigeração e sanitização para garantir segurança.This study evaluated a biological control system for the inhibition of Listeria monocytogenes in minimally processed kale focusing on its freshness under refrigeration and extreme temperatures. The inhibitory potential of salt and cold tolerant lactic bacteria from natural microflora of kale, Lactobacillus delbrueckii ATCC 9649, Lactobacillus plantarum, and Lactobacillus casei CCT 1465 strains were evaluated against L. monocytogenes. Pediococcus acidilactici CCA3 isolated from kale exhibited a large inhibition zone of L. monocytogenes at 10 and 15 °C in MRS agar and was

  14. The major volatile compound 2-phenylethanol from the biocontrol yeast, Pichia anomala, inhibits growth and expression of aflatoxin biosynthetic genes of Aspergillus flavus.

    Science.gov (United States)

    Hua, Sui Sheng T; Beck, John J; Sarreal, Siov Bouy L; Gee, Wai

    2014-05-01

    Aspergillus flavus is a ubiquitous saprophyte that is able to produce the most potent natural carcinogenic compound known as aflatoxin B1 (AFB1). This toxin frequently contaminates crops including corn, cotton, peanuts, and tree nuts causing substantial economic loss worldwide. Consequently, more than 100 countries have strict regulations limiting AFB1 in foodstuffs and feedstuffs. Plants and microbes are able to produce volatile compounds that act as a defense mechanism against other organisms. Pichia anomala strain WRL-076 is a biocontrol yeast currently being tested to reduce AF contamination of tree nuts in California. We used the SPME-GC/MS analysis and identified the major volatile compound produced by this strain to be 2-phenylethanol (2-PE). It inhibited spore germination and AF production of A. flavus. Inhibition of AF formation by 2-PE was correlated with significant down regulation of clustering AF biosynthesis genes as evidenced by several to greater than 10,000-fold decrease in gene expression. In a time-course analysis we found that 2-PE also altered the expression patterns of chromatin modifying genes, MYST1, MYST2, MYST3, gcn5, hdaA and rpdA. The biocontrol capacity of P. anomala can be attributed to the production of 2-PE, which affects spore germination, growth, toxin production, and gene expression in A. flavus. PMID:24504634

  15. The effect of locust bean gum (LBG)-based edible coatings carrying biocontrol yeasts against Penicillium digitatum and Penicillium italicum causal agents of postharvest decay of mandarin fruit.

    Science.gov (United States)

    Parafati, Lucia; Vitale, Alessandro; Restuccia, Cristina; Cirvilleri, Gabriella

    2016-09-01

    Strains belonging to Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans species were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic molds Penicillium digitatum and Penicillium italicum. Moreover, studies aimed at screening the antifungal activity of selected yeast strains in vivo conditions against P. digitatum and P. italicum, and investigated the efficacy of a polysaccharidic matrix, locust bean gum (LBG), enriched with the tested BCAs, in controlling postharvest decays in artificially inoculated mandarins. The population dynamics of BCAs on wounds and the magnitude of peroxidase (POD) and superoxide dismutase (SOD) in fruit tissues were also investigated after treatments of mandarins with antagonistic yeasts. W. anomalus BS91, M. pulcherrima MPR3 and A. pullulans PI1 provided excellent control of postharvest decays caused by P. digitatum and P. italicum on mandarins, both when the yeasts were used alone and in combination with LBG, which enhanced the yeast cell viability over time. Finally, the increased activity of POD and lower decrease in SOD activity in response to BCAs application in mandarin fruits confirmed their involvement in the biocontrol mechanism. PMID:27217363

  16. Evaluation of the atoxigenic Aspergillus flavus strain AF36 in pistachio orchards

    Science.gov (United States)

    The atoxigenic strain Aspergillus flavus AF36, which has been extensively used as a biocontrol agent in commercial corn and cotton fields to reduce aflatoxin contamination, was applied in research pistachio orchards from 2002 to 2005 and in commercial pistachio orchards from 2008 to 2011. AF36 was a...

  17. The Sensor Kinase GacS Negatively Regulates Flagellar Formation and Motility in a Biocontrol Bacterium, Pseudomonas chlororaphis O6

    Directory of Open Access Journals (Sweden)

    Ji Soo Kim

    2014-06-01

    Full Text Available The GacS/GacA two component system regulates various traits related to the biocontrol potential of plant-associated pseudomonads. The role of the sensor kinase, GacS, differs between strains in regulation of motility. In this study, we determined how a gacS mutation changed cell morphology and motility in Pseudomonas chlororaphis O6. The gacS mutant cells were elongated in stationary-phase compared to the wild type and the complemented gacS mutant, but cells did not differ in length in logarithmic phase. The gacS mutant had a two-fold increase in the number of flagella compared with the wild type strain; flagella number was restored to that of the wild type in the complemented gacS mutant. The more highly flagellated gacS mutant cells had greater swimming motilities than that of the wild type strain. Enhanced flagella formation in the gacS mutant correlated with increased expression of three genes, fleQ, fliQ and flhF, involved in flagellar formation. Expression of these genes in the complemented gacS mutant was similar to that of the wild type. These findings show that this root-colonizing pseudomonad adjusts flagella formation and cell morphology in stationary-phase using GacS as a major regulator.

  18. Biocontrol of Postharvest Anthracnose of Mango Fruit with Debaryomyces Nepalensis and Effects on Storage Quality and Postharvest Physiology.

    Science.gov (United States)

    Luo, Shanshan; Wan, Bin; Feng, Shuhan; Shao, Yuanzhi

    2015-11-01

    Anthracnose is presently recognized as one of the most important postharvest disease of mango worldwide. To control the disease, chemical fungicides for a long time was widely used among fruit farmers, but recently found that pathogen had developed increasingly resistance to it. With people's growing desire of healthy and green food, finding new and environmentally friendly biological control approach was very necessary. In this paper, we provided a kind of new antagonistic yeast which enriched the strain resources and the efficacy of Debaryomyces nepalensis against postharvest anthracnose of mango fruit and the influence on quality parameters were investigated. The results showed that the decay incidence and lesion diameter of postharvest anthracnose of mango treated by D. nepalensis were significantly reduced compared with the control fruit stored at 25 °C for 30 d or at 15 °C for 40 d, and the higher concentration of D. nepalensis was, the better the efficacy of the biocontrol was. Study also found that 1 h was the best treatment duration and antagonistic yeast inoculated earlier had good biocontrol effect on anthracnose. Meanwhile, treatment by D. nepalensis could significantly reduce postharvest anthracnose of mango, delay the decrease in firmness, TSS, TA, and ascorbic acid value, and do not impair surface color during postharvest storage. Moreover, the increase in MDA (malondialdehyde) content and increase in cell membrane permeability of fruit treated by D. nepalensis was highly inhibited. The results suggested D. nepalensis treatment could not only maintain storage quality of mango fruit, but also decrease the decay incidence to anthracnose disease. All these results indicated that D. nepalensis has great potential for development of commercial formulations to control postharvest pathogens of mango fruit.

  19. Whole-Genome Shotgun Sequence of Bacillus mojavensis Strain RRC101, an Endophytic Bacterium Antagonistic to the Mycotoxigenic Endophytic Fungus Fusrium verticillioides

    Science.gov (United States)

    Here we report the whole genome shotgun sequence of Bacillus mojavensis strain RRC101, isolated from a maize kernel. This strain is antagonistic to the mycotoxigenic plant pathogen Fusarium verticillioides, and grows within maize tissue, suggesting potential as an endophytic biocontrol agent....

  20. Whole-Genome Shotgun Sequence of Bacillus mojavensis Strain RRC101, an Endophytic Bacterium Antagonistic to the Mycotoxigenic Endophytic Fungus Fusarium verticillioides

    OpenAIRE

    Gold, S. E.; Blacutt, A. A.; Meinersmann, R. J.; Bacon, C W

    2014-01-01

    Here, we report the whole-genome shotgun sequence of Bacillus mojavensis strain RRC101, isolated from a maize kernel. This strain is antagonistic to the mycotoxigenic plant pathogen Fusarium verticillioides and grows within maize tissue, suggesting potential as an endophytic biocontrol agent.

  1. Identification of a NovelPaenibacillus polymyxa Strain and Its Biocontrol and Plant Growth-Promoting Effects%一株多粘类芽孢杆菌的鉴定及其生防促生效果初步测定

    Institute of Scientific and Technical Information of China (English)

    郭芳芳; 谢镇; 卢鹏; 郭岩彬; 张立钦; 王勇军

    2014-01-01

    菌株CF05是从浙江天目山柳杉中分离得到的1株细菌,经形态、16S rDNA序列、生理生化指标检测,鉴定为多粘类芽孢杆菌Paenibacillus polymyxa。平板拮抗结果表明,该菌对9种植物病原真菌和3种病原细菌具有显著拮抗效果。室内生测结果表明,发酵液和菌悬液对番茄幼苗均具有明显的促生效果,其中发酵液处理最佳,鲜重增长272.0%,干重增长266.7%。菌株CF05接种27 d后,番茄根际细菌浓度仍很高。种子萌发测定结果表明,菌株 CF05处理后发芽率提高11.7%,番茄猝倒病防治测定结果显示,发病率降低38.6%,效果均优于Bacillus subtilis 168处理。综上结果,表明P. polymyxa CF05是1株生防性状优良的促生细菌。%Strain CF05 isolated from Cryptomeria fortunei in Tianmu Mountain, Zhejiang province was identified asPaenibacillus polymyxa on the bases of its phenotypical and biochemical characteristics as well as 16S rDNA gene sequence. TheP. polymyxaCF05 exhibited highly antagonistic effects against 9 plant fungal pathogens and 3 plant bacterial pathogensin vitro. Furthermore, both cell-free fermentation and bacterial suspension promoted tomato seedlings growth in greenhouse assay. Cell-free fermentation increased the biomass with fresh weight and dry weight by 272.0% and 266.7%, respectively. It was demonstrated that secondary metabolites from this bacterium might perform a critical function in plant growth promotion. Tests of root-colonizing ability indicated that concentrations of the bacterium in tomato rhizosphere were 3.8, 3.6, 3.1 and 3.1 log CFU/g root after 27 days of inoculation of tomato seeds with the bacterial suspension at 108、107、106、105 CFU/mL, respectively. Seed germination ratio was increased by 11.7%, and the disease incidence of damping-off was reduced by 38.6% after seed treatment.

  2. The SRAP based molecular diversity related to antifungal and antioxidant bioactive constituents for biocontrol potentials of Trichoderma against Sclerotium rolfsii Scc.

    Science.gov (United States)

    Hirpara, Darshna G; Gajera, H P; Bhimani, R D; Golakiya, B A

    2016-08-01

    The study was performed to examine 11 isolates of Trichoderma for their bio-control potentials against Sclerotium rolfsii Sacc. causing stem rot in groundnut. The antagonists Trichoderma were subjected to sequence related amplified polymorphism (SRAP) based molecular diversity analysis and compared with their hardness to S. rolfsii with respect to secretary antifungal and antioxidant profile. T. virens NBAII Tvs 12 evident highest (87.91 %) growth inhibition of test pathogen followed by T. koningii MTCC 796 (67.03 %) at 7 days after inoculation (DAI). Microscopic study confirmed biocontrol mechanism as mycoparasitism for Tvs 12 and antibiosis for MTCC 796. The growth inhibition of test pathogen was significantly negatively correlated with sclerotia formation and lipid peroxidation during antagonism due to release of secretary bioactive antioxidants by antagonists to terminate oxidative burst generated by S. rolfsii and causing inhibition of sclerotium formation. The GC-MS profile identified antifungal and antioxidant constituents hexadecane, 1,2-benzenedicarboxylic acid, mono (2-ethylhexyl) ester, 1-hexadecanesulfonyl chloride, and octadecane in potent antagonists Tvs 12; and nonacosane and octadecane in MTCC 796 along with two novel compounds 1-pentadecene and 1-heneicosyl formate for biocontrol activity. Molecular diversity of Trichoderma isolates associated with antagonistic activity was assessed by SRAP markers. The 115 primer combinations generate total 1328 amplified products of which, 1095 are shared polymorphic and 199 are unique polymorphic. The 15 SRAP combinations produced 18 bands to diagnose best antagonist Tvs 12 and 13 SRAP combinations generated 19 unique bands for identification of MTCC 796. The mycoparasitic antagonist Tvs 12 would be the best antagonist and released unique antifungal and antioxidant constituents to combat pathogen infection. The SRAP based genetic diversity indicates Tvs12 strain clustered with T. viride NBAII Tv23 and shared

  3. Biocontrol of Botrytis cinerea by successful introduction of Pantoea ananatis in the grapevine phyllosphere

    Directory of Open Access Journals (Sweden)

    Gasser F

    2012-12-01

    Full Text Available Florian Gasser,1 Massimiliano Cardinale,1 Barbara Schildberger,2 Gabriele Berg11Institute of Environmental Biotechnology, Graz University of Technology, Graz, Austria; 2Höhere Bundesanstalt und Bundesamt für Wein-und Obstbau, Klosterneuburg, AustriaBackground and aims: The fungus Botrytis cinerea is a common problem in viticulture and leads to serious losses in both yield and quality. The objective was to study the potential of the antagonist Pantoea ananatis BLBT1-08 for controlling this disease.Methods: Pathogen suppression by Pantoea treatments was investigated in different field trials and in detached leaf assays. The mode of action was studied by confocal laser scanning microscopy of treated grape leaves and by in vitro assays.Results: The introduction of P. ananatis BLBT1-08 in a 3-year field trial resulted in statistically significant reduction of disease symptoms. However, B. cinerea abundance, measured by quantitative real-time polymerase chain reaction of a B. cinerea specific gene, was not reduced when compared to non-treated, symptom-free leaves. A DsRed fluorescent protein labeled BLBT1-08 strain showed a high phyllosphere competence and competition on the leaf surface, but did not colonize the inner parts of plant tissue. Germination of B. cinerea was not inhibited by BLBT1-08 on the leaf, but mycelial growth and symptoms were suppressed without direct pathogen-antagonist contact. The antimicrobial activity was amino acid and temperature dependent.Conclusion: P. ananatis BLBT1-08 is a competitive and promising biocontrol agent for the control of B. cinerea and is highly effective at reducing disease incidence.Keywords: biological control, sustainable viticulture, antagonism

  4. Use of AFLP for differentiation of Metschnikowia pulcherrima strains for postharvest disease biological control.

    Science.gov (United States)

    Spadaro, D; Sabetta, W; Acquadro, A; Portis, E; Garibaldi, A; Gullino, M L

    2008-01-01

    Metschnikowia pulcherrima occurs naturally on fruits, buds and floral parts of apple trees. Some strains are effective as biocontrol agents against postharvest decay of apples and other fruits. The usefulness of the amplified fragment length polymorphism (AFLP) technique was evaluated for the genetic analysis of 26 strains of M. pulcherrima, isolated from different sources in different geographical regions. With six AFLP primer pairs, 729 polymorphic bands were scored. The technique showed a high discriminatory power. Genetic relationships between strains were also estimated using AFLP. All the isolates from the carposphere of apple, previously tested as biocontrol agents, were grouped in a single cluster with a high bootstrap value (97), indicating robustness and reproducibility. AFLP patterns could clearly distinguish the different strains and research is in progress to use some putative specific bands for single tag sequence (STS) conversion to develop isolate-specific markers.

  5. Utilization of winery wastes for Trichoderma viride biocontrol agent production by solid state fermentation

    Institute of Scientific and Technical Information of China (English)

    BAI Zhihui; JIN Bo; LI Yuejie; CHEN Jian; LI Zuming

    2008-01-01

    Biocontrol agents are safe and environmental friendly alternatives for pesticides in agriculture application.Trichoderma v/ride WEBL0703 performed a high level of antagonistic activity toward a broad spectrum of phytopathogens and was determined as a biocontrol agent,which was produced by solid state fermentation using grape marc and wine lees.The maximum yield of T.viride conidia was up to 6.65×109 CFU/g initial dry substrate (IDS) after 10 d fermentation.As important enzymes for protecting plants from disease,ehitinase,β-glucanase,and pectinase yields were 47.8 U/g IDS,8.32 U/g IDS and 9.83 U/g IDS,respectively.These results show that it is feasible to convert winery wastes to a value-added and environmental friendly biocontrol agent.

  6. Antibiosis by Pantoea agglomerans biocontrol strain E325 against Erwinia amylovora on apple blossom stigmas

    Science.gov (United States)

    Pantoea agglomerans E325, the active ingredient in a commercial product for fire blight control, was previously shown in vitro to produce a unique alkaline- and phosphate-sensitive antibiotic specific to Erwinia amylovora. Antibiosis was evaluated as a mode of antagonism on blossom stigmas using two...

  7. Complete genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium of Calendula officinalis

    Energy Technology Data Exchange (ETDEWEB)

    Koeberl, Martina; White, Richard A.; Erschen, Sabine; Spanberger, Nora; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-08-13

    The genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium (PGPR) with broad-spectrum antagonistic activities against plant pathogenic fungi, bacteria and nematodes, consists of a single 3.9 Mb circular chromosome. The genome reveals genes putatively responsible for its promising biocontrol and PGP properties.

  8. Complete Genome Sequence of Bacillus amyloliquefaciens Strain Co1-6, a Plant Growth-Promoting Rhizobacterium of Calendula officinalis.

    Science.gov (United States)

    Köberl, Martina; White, Richard A; Erschen, Sabine; Spanberger, Nora; El-Arabi, Tarek F; Jansson, Janet K; Berg, Gabriele

    2015-01-01

    The genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium (PGPR) with broad-spectrum antagonistic activity against plant-pathogenic fungi, bacteria, and nematodes, consists of a single 3.9-Mb circular chromosome. The genome reveals genes putatively responsible for its promising biocontrol and PGP properties. PMID:26272562

  9. Complete Genome Sequence of Bacillus amyloliquefaciens Strain Co1-6, a Plant Growth-Promoting Rhizobacterium of Calendula officinalis

    OpenAIRE

    Köberl, Martina; White, Richard A.; Erschen, Sabine; Spanberger, Nora; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-01-01

    The genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium (PGPR) with broad-spectrum antagonistic activity against plant-pathogenic fungi, bacteria, and nematodes, consists of a single 3.9-Mb circular chromosome. The genome reveals genes putatively responsible for its promising biocontrol and PGP properties.

  10. Biosynthetic mechanism for sunscreens of the biocontrol agent Lysobacter enzymogenes.

    Directory of Open Access Journals (Sweden)

    Yan Wang

    Full Text Available Lysobacter are ubiquitous environmental bacteria emerging as novel biocontrol agents and new sources of anti-infectives. So far, very little effort has been invested in the study of the biology of these Gram-negative gliding bacteria. Many Lysobacter species are characterized by their yellow-orange appearance. Using transposon mutagenesis, we identified a stand-alone polyketide synthase (PKS gene cluster required for the pigment production in L. enzymogenes OH11. The yellow pigments were abolished in the "white" mutants generated by target-specific deletions of ketosynthase (KS, acyl carrier protein, or ketoreductase. Spectroscopic data suggested that the pigments belong to xanthomonadin-like aryl polyenes. Polyene-type polyketides are known to be biosynthesized by modular PKS (Type I, not by stand-alone PKS (Type II which always contain the heterodimer KS-CLF (chain-length factor as the key catalytic component. Remarkably, this aryl polyene PKS complex only contains the KS (ORF17, but not the CLF. Instead, a hypothetical protein (ORF16 is located immediately next to ORF17. ORF16-17 homologs are widespread in numerous uncharacterized microbial genomes, in which an ORF17 homolog is always accompanied by an ORF16 homolog. The deletion of ORF16 eliminated pigment production, and homology modeling suggested that ORF16 shares a structural similarity to the N-terminal half of CLF. A point-mutation of glutamine (Q166A that is the conserved active site of known CLF abolished pigment production. The "white" mutants are significantly more sensitive to UV/visible light radiation or H2O2 treatment than the wild type. These results unveil the first example of Type II PKS-synthesized polyene pigments and show that the metabolites serve as Lysobacter "sunscreens" that are important for the survival of these ubiquitous environmental organisms.

  11. Tick control: trapping, biocontrol, host management and other alternative strategies

    Science.gov (United States)

    Ginsberg, Howard S.; Edited by Sonenshine, Daniel E.; Roe, R. Michael

    2014-01-01

    Biology of Ticks is the most comprehensive work on tick biology and tick-borne diseases. This second edition is a multi-authored work, featuring the research and analyses of renowned experts across the globe. Spanning two volumes, the book examines the systematics, biology, structure, ecological adaptations, evolution, genomics and the molecular processes that underpin the growth, development and survival of these important disease-transmitting parasites. Also discussed is the remarkable array of diseases transmitted (or caused) by ticks, as well as modern methods for their control. This book should serve as a modern reference for students, scientists, physicians, veterinarians and other specialists. Volume I covers the biology of the tick and features chapters on tick systematics, tick life cycles, external and internal anatomy, and others dedicated to specific organ systems, specifically, the tick integument, mouthparts and digestive system, salivary glands, waste removal, salivary glands, respiratory system, circulatory system and hemolymph, fat body, the nervous and sensory systems and reproductive systems. Volume II includes chapters on the ecology of non-nidicolous and nidicolous ticks, genetics and genomics (including the genome of the Lyme disease vector Ixodes scapularis) and immunity, including host immune responses to tick feeding and tick-host interactions, as well as the tick's innate immune system that prevents and/or controls microbial infections. Six chapters cover in depth the many diseases caused by the major tick-borne pathogens, including tick-borne protozoa, viruses, rickettsiae of all types, other types of bacteria (e.g., the Lyme disease agent) and diseases related to tick paralytic agents and toxins. The remaining chapters are devoted to tick control using vaccines, acaricides, repellents, biocontrol, and, finally, techniques for breeding ticks in order to develop tick colonies for scientific study.

  12. Cytokinin production by Pseudomonas fluorescens G20-18 determines biocontrol activity against Pseudomonas syringae in Arabidopsis.

    Science.gov (United States)

    Großkinsky, Dominik K; Tafner, Richard; Moreno, María V; Stenglein, Sebastian A; García de Salamone, Inés E; Nelson, Louise M; Novák, Ondřej; Strnad, Miroslav; van der Graaff, Eric; Roitsch, Thomas

    2016-01-01

    Plant beneficial microbes mediate biocontrol of diseases by interfering with pathogens or via strengthening the host. Although phytohormones, including cytokinins, are known to regulate plant development and physiology as well as plant immunity, their production by microorganisms has not been considered as a biocontrol mechanism. Here we identify the ability of Pseudomonas fluorescens G20-18 to efficiently control P. syringae infection in Arabidopsis, allowing maintenance of tissue integrity and ultimately biomass yield. Microbial cytokinin production was identified as a key determinant for this biocontrol effect on the hemibiotrophic bacterial pathogen. While cytokinin-deficient loss-of-function mutants of G20-18 exhibit impaired biocontrol, functional complementation with cytokinin biosynthetic genes restores cytokinin-mediated biocontrol, which is correlated with differential cytokinin levels in planta. Arabidopsis mutant analyses revealed the necessity of functional plant cytokinin perception and salicylic acid-dependent defence signalling for this biocontrol mechanism. These results demonstrate microbial cytokinin production as a novel microbe-based, hormone-mediated concept of biocontrol. This mechanism provides a basis to potentially develop novel, integrated plant protection strategies combining promotion of growth, a favourable physiological status and activation of fine-tuned direct defence and abiotic stress resilience. PMID:26984671

  13. Yeast Biocontrol of a Fungal Plant Disease: A Model for Studying Organism Interrelationships

    Science.gov (United States)

    Chanchaichaovivat, Arun; Panijpan, Bhinyo; Ruenwongsa, Pintip

    2008-01-01

    An experiment on the action of the yeast, "Saccharomyces cerevisiae", against a fungal plant disease is proposed for secondary students (Grade 11) to support their study of organism interrelationship. This biocontrol experiment serves as the basis for discussing relationships among three organisms (red chilli fruit, "Saccharomyces cerevisiae," and…

  14. What magnitude are observed non-target impacts from weed biocontrol?

    Science.gov (United States)

    A systematic review focused by plant on non-target impacts from agents deliberately introduced for the biological control of weeds found significant non-target impacts to be rare. The magnitude of direct impact of 43 biocontrol agents on 140 non-target plants was retrospectively categorized using a ...

  15. Agar composition affects in vitro screening of biocontrol activity of antagonistic microorganisms

    NARCIS (Netherlands)

    Bosmans, Lien; De Bruijn, I.; de Mot, Rene; Readers, Hans; Lievens, Bart

    2016-01-01

    Agar-based screening assays are the method of choice when evaluating antagonistic potential of bacterial biocontrol-candidates against pathogens.Weshowed thatwhen using the samemedium, but different agar compositions, the activity of a bacterial antagonist against Agrobacteriumwas strongly affected.

  16. Stress tolerance and biocontrol performance of the yeast antagonist, Candida diversa, change with morphology transition.

    Science.gov (United States)

    Li, Guangkun; Chi, Mengshan; Chen, Huizhen; Sui, Yuan; Li, Yan; Liu, Yongsheng; Zhang, Xiaojing; Sun, Zhiqiang; Liu, Guoqing; Wang, Qi; Liu, Jia

    2016-02-01

    As an eco-friendly management method, biological control of postharvest diseases, utilizing antagonistic yeasts, is a research topic receiving considerable attention. Detailed knowledge on the biology of yeast antagonists is crucial when considering their potential application and development as biocontrol products. Changes in the growth form, such as single-cell to pseudohyphae, have been associated with the mode of action in postharvest biocontrol yeasts. In this study, the antagonistic yeast, Candida diversa, reversibly shifted from a single-cell morphology on yeast peptone dextrose (YPD) medium with 2 % agar to a pseudohyphal morphology on YPD with 0.3 % agar. The tolerance of the pseudohyphal form to heat and oxidative stresses, as well as the biocontrol efficacy against Botrytis cinerea on apple and kiwifruit stored at 25 and 4 °C, was significantly higher as compared to the single-cell form. This study provides new information on the ability of C. diversa to change its morphology and the impact of the morphology shift on stress tolerance and biocontrol performance.

  17. Draft Whole-Genome Sequence of the Biocontrol Agent Trichoderma harzianum T6776.

    Science.gov (United States)

    Baroncelli, Riccardo; Piaggeschi, Giulia; Fiorini, Lisa; Bertolini, Edoardo; Zapparata, Antonio; Pè, Mario Enrico; Sarrocco, Sabrina; Vannacci, Giovanni

    2015-01-01

    Trichoderma harzianum T6776 is a promising beneficial isolate whose effects consist of growth promotion, positive response of photosynthetic activity, hormonal signaling, and carbon partitioning in tomato, coupled with biocontrol of plant pathogens. Here, we present the first genome assembly of T6776, providing a useful platform for the scientific community. PMID:26067977

  18. Biocontrol potential of endophytes harbored in Radula marginata (liverwort) from the New Zealand ecosystem.

    Science.gov (United States)

    Kusari, Parijat; Kusari, Souvik; Spiteller, Michael; Kayser, Oliver

    2014-10-01

    Radula marginata and Cannabis sativa L. are two phylogenetically unrelated plant species containing structurally similar secondary metabolites like cannabinoids. The major objective of our work was the isolation, identification, biocontrol efficacies, biofilm forming potential and anti-biofilm ability of endophytic microbial community of the liverwort R. marginata, as compared to bacterial endophytic isolates harbored in C. sativa plants. A total of 15 endophytic fungal and 4 endophytic bacterial isolates were identified, including the presence of a bacterial endosymbiont within an endophytic fungal isolate. The endosymbiont was visible only when the fungus containing it was challenged with two phytopathogens Botrytis cinerea and Trichothecium roseum, highlighting a tripartite microbe-microbe interaction and biocontrol potency of endophytes under biotic stress. We also observed sixteen types of endophytic fungal-pathogen and twelve types of endophytic bacterial-pathogen interactions coupled to varying degree of growth inhibitions of either the pathogen or endophyte or both. This showed the magnitude of biocontrol efficacies of endophytes in aiding plant fitness benefits under different media (environmental) conditions. Additionally, it was ecologically noteworthy to find the presence of similar endophytic bacterial genera in both Radula and Cannabis plants, which exhibited similar functional traits like biofilm formation and general anti-biofilm activities. Thus far, our work underlines the biocontrol potency and defensive functional traits (in terms of antagonism and biofilm formation) of endophytes harbored in liverwort R. marginata as compared to the endophytic community of phylogenetically unrelated but phytochemically similar plant C. sativa. PMID:25100187

  19. Stress tolerance and biocontrol performance of the yeast antagonist, Candida diversa, change with morphology transition.

    Science.gov (United States)

    Li, Guangkun; Chi, Mengshan; Chen, Huizhen; Sui, Yuan; Li, Yan; Liu, Yongsheng; Zhang, Xiaojing; Sun, Zhiqiang; Liu, Guoqing; Wang, Qi; Liu, Jia

    2016-02-01

    As an eco-friendly management method, biological control of postharvest diseases, utilizing antagonistic yeasts, is a research topic receiving considerable attention. Detailed knowledge on the biology of yeast antagonists is crucial when considering their potential application and development as biocontrol products. Changes in the growth form, such as single-cell to pseudohyphae, have been associated with the mode of action in postharvest biocontrol yeasts. In this study, the antagonistic yeast, Candida diversa, reversibly shifted from a single-cell morphology on yeast peptone dextrose (YPD) medium with 2 % agar to a pseudohyphal morphology on YPD with 0.3 % agar. The tolerance of the pseudohyphal form to heat and oxidative stresses, as well as the biocontrol efficacy against Botrytis cinerea on apple and kiwifruit stored at 25 and 4 °C, was significantly higher as compared to the single-cell form. This study provides new information on the ability of C. diversa to change its morphology and the impact of the morphology shift on stress tolerance and biocontrol performance. PMID:26637302

  20. Incorporating Ecologically Relevant Measures of Pesticide Effect for Estimating the Compatibility of Pesticides and Biocontrol Agents

    Science.gov (United States)

    The compatibility of biological control agents with pesticides is a central concern in integrated pest management programs. The most common assessments of compatibility consist of simple comparisons of acute toxicity among pest species and select biocontrol agents. A more sophisticated approach, d...

  1. Isolation and identification of actinomycetes from a compost-amended soils biocontrol agents

    Energy Technology Data Exchange (ETDEWEB)

    Garcia de la Fuente, R.; Cuesta, G.; Fornes, F.; Abad, M.

    2009-07-01

    Compost capability to suppress soil-borne plant pathogens has become an interesting subject as a strategy for reducing the adverse effects of massive fungicides application in the environmental. In this context, actinomycetes have received considerable attention as biocontrol agents, particularly Streptomyces species. (Author)

  2. The ability of a cold-adapted Rhodotorula mucilaginosa strain from Tibet to control blue mold in pear fruit.

    Science.gov (United States)

    Hu, Hao; Yan, Fujie; Wilson, Charles; Shen, Qing; Zheng, Xiaodong

    2015-12-01

    Cold-adapted yeasts were isolated from soil samples collected in Tibet and evaluated as potential biocontrol agents against blue mold (Penicillium expansum) of pear fruit in cold storage. YC1, an isolate identified as Rhodotorula mucilaginosa, was found to exhibit the greatest biocontrol activity among the different isolates that were screened. A washed cell suspension of YC1 exhibited the best biocontrol activity among three different preparations that were used in the current study. A concentration of 10(8) cells/ml reduced the incidence of decay to 35 %, compared to the control where decay incidence was 100 %. A higher intracellular level of trehalose and a higher proportion of polyunsaturated acids present in YC1, was associated with increased the tolerance of this strain to low temperatures, relative to the other strains that were evaluated. The increased tolerance to low temperature allowed the YC1 strain of yeast to more effectively compete for nutrients and space in wounded pear fruit that had been inoculated with spores of P. expansum and placed in cold storage. The present study demonstrated the ability to select cold-adapted yeasts from cold climates and use them as biocontrol agents of postharvest diseases of fruit placed in cold storage. PMID:26454432

  3. Biocontrol of Late Blight (Phytophthora capsici Disease and Growth Promotion of Pepper by Burkholderia cepacia MPC-7

    Directory of Open Access Journals (Sweden)

    Mao Sopheareth

    2013-03-01

    Full Text Available A chitinolytic bacterial strain having strong antifungal activity was isolated and identified as Burkholderia cepacia MPC-7 based on 16S rRNA gene analysis. MPC-7 solubilized insoluble phosphorous in hydroxyapatite agar media. It produced gluconic acid and 2-ketogluconic acid related to the decrease in pH of broth culture. The antagonist produced benzoic acid (BA and phenylacetic acid (PA. The authentic compounds, BA and PA, showed a broad spectrum of antimicrobial activity against yeast, several bacterial and fungal pathogens in vitro. To demonstrate the biocontrol efficiency of MPC-7 on late blight disease caused by Phytophthora capsici, pepper plants in pot trials were treated with modified medium only (M, M plus zoospore inoculation (MP, MPC-7 cultured broth (B and B plus zoospore inoculation (BP. With the sudden increase in root mortality, plants in MP wilted as early as five days after pathogen inoculation. However, plant in BP did not show any symptom of wilting until five days. Root mortality in BP was markedly reduced for as much as 50%. Plants in B had higher dry weight, P concentration in root, and larger leaf area compared to those in M and MP. These results suggested that B. cepacia MPC-7 should be considered as a candidate for the biological fertilizer as well as antimicrobial agent for pepper plants.

  4. Trichoderma Biocontrol: Signal Transduction Pathways Involved in Host Sensing and Mycoparasitism

    Directory of Open Access Journals (Sweden)

    Susanne Zeilinger

    2007-01-01

    Full Text Available Fungi of the genus Trichoderma are used as biocontrol agents against several plant pathogenic fungi like Rhizoctonia spp., Pythium spp., Botrytis cinerea and Fusarium spp. which cause both soil-borne and leaf- or flower-borne diseases of agricultural plants. Plant disease control by Trichoderma is based on complex interactions between Trichoderma, the plant pathogen and the plant. Until now, two main components of biocontrol have been identified: direct activity of Trichoderma against the plant pathogen by mycoparasitism and induced systemic resistance in plants. As the mycoparasitic interaction is host-specific and not merely a contact response, it is likely that signals from the host fungus are recognised by Trichoderma and provoke transcription of mycoparasitism-related genes.In the last few years examination of signalling pathways underlying Trichoderma biocontrol started and it was shown that heterotrimeric G-proteins and mitogen-activated protein (MAP kinases affected biocontrol-relevant processes such as the production of hydrolytic enzymes and antifungal metabolites and the formation of infection structures. MAPK signalling was also found to be involved in induction of plant systemic resistance in Trichoderma virens and in the hyperosmotic stress response in Trichoderma harzianum. Analyses of the function of components of the cAMP pathway during Trichoderma biocontrol revealed that mycoparasitism-associated coiling and chitinase production as well as secondary metabolism are affected by the internal cAMP level; in addition, a cross talk between regulation of light responses and the cAMP signalling pathway was found in Trichoderma atroviride.

  5. Biocontrol potential of Trichoderma harzianum isolate T-aloe against Sclerotinia sclerotiorum in soybean.

    Science.gov (United States)

    Zhang, Fuli; Ge, Honglian; Zhang, Fan; Guo, Ning; Wang, Yucheng; Chen, Long; Ji, Xiue; Li, Chengwei

    2016-03-01

    Sclerotinia stem rot, caused by Sclerotinia sclerotiorum (Lib.) de Bary is a major disease of soybean (Glycine max (L.) Merr.). At present, we revealed the three-way interaction between Trichoderma harzianum T-aloe, pathogen S. sclerotiorum and soybean plants in order to demonstrate biocontrol mechanism and evaluate biocontrol potential of T-aloe against S. sclerotiorum in soybean. In our experiments, T-aloe inhibited the growth of S. sclerotiorum with an efficiency of 56.3% in dual culture tests. T-aloe hyphae grew in parallel or intertwined with S. sclerotiorum hyphae and produced hooked contact branches, indicating mycoparasitism. Plate tests showed that T-aloe culture filtrate inhibited S. sclerotiorum growth with an inhibition efficiency of 51.2% and sclerotia production. T-aloe pretreatment showed growth-promoting effect on soybean plants. The activities of peroxidase, superoxide dismutase, and catalase increased, and the hydrogen peroxide (H2O2) as well as the superoxide radical (O2(-)) content in soybean leaves decreased after T-aloe pretreatment in response to S. sclerotiorum pathogen challenge. T-aloe treatment diminished damage caused by pathogen stress on soybean leaf cell membrane, and increased chlorophyll as well as total phenol contents. The defense-related genes PR1, PR2, and PR3 were expressed in the leaves of T-aloe-treated plants. In summary, T-aloe displayed biocontrol potential against S. sclerotiorum. This is the first report of unraveling biocontrol potential of Trichoderma Spp. to soybean sclerotinia stem rot from the three-way interaction between the biocontrol agent, pathogen S. sclerotiorum and soybean plants. PMID:26774866

  6. Selection of the N-acylhomoserine lactone-degrading bacterium Alteromonas stellipolaris PQQ-42 and of its potential for biocontrol in aquaculture

    Directory of Open Access Journals (Sweden)

    Marta eTorres

    2016-05-01

    Full Text Available The production of virulence factors by many pathogenic microorganisms depends on the intercellular communication system called quorum sensing (QS, which involves the production and release of signal molecules known as autoinducers. Based on this, new-therapeutic strategies have emerged for the treatment of a variety of infections, such as the enzymatic degradation of signalling molecules, known as quorum quenching (QQ. In this study, we present the screening of QQ activity amongst 450 strains isolated from a bivalve hatchery in Granada (Spain, and the selection of the strain PQQ-42, which degrades a wide range of N-acylhomoserine lactones (AHLs. The selected strain, identified as Alteromonas stellipolaris, degraded the accumulation of AHLs and reduced the production of protease and chitinase and swimming motility of a Vibrio species in co-cultivation experiments in vitro. In the bio-control experiment, strain PQQ-42 significantly reduced the pathogenicity of V. mediterranei VibC-Oc-097 upon the coral Oculina patagonica showing a lower degree of tissue damage (29.25±14.63 % in its presence, compared to when the coral was infected with V. mediterranei VibC-Oc-097 alone (77.53±13.22 %. Our results suggest that this AHL-degrading bacterium may have biotechnological applications in aquaculture.

  7. Evaluation of the sea anemone Anthothoe albocincta as an augmentative biocontrol agent for biofouling on artificial structures.

    Science.gov (United States)

    Atalah, Javier; Bennett, Holly; Hopkins, Grant A; Forrest, Barrie M

    2013-01-01

    Augmentative biocontrol, defined as the use of indigenous natural enemies to control pest populations, has not been explored extensively in marine systems. This study tested the potential of the anemone Anthothoe albocincta as a biocontrol agent for biofouling on submerged artificial structures. Biofouling biomass was negatively related to anemone cover. Treatments with high anemone cover (>35%) led to significant changes in biofouling assemblages compared to controls. Taxa that contributed to these changes differed among sites, but included reductions in cover of problematic fouling organisms, such as solitary ascidians and bryozoans. In laboratory trials, A. albocincta substantially prevented the settlement of larvae of the bryozoan Bugula neritina when exposed to three levels of larval dose, suggesting predation as an important biocontrol mechanism, in addition to space pre-emption. This study demonstrated that augmentative biocontrol using anemones has the potential to reduce biofouling on marine artificial structures, although considerable further work is required to refine this tool before its application.

  8. Draft genome sequence of Streptomyces sp. strain Wb2n-11, a desert isolate with broad-spectrum antagonism against soilborne phytopathogens

    Energy Technology Data Exchange (ETDEWEB)

    Koeberl, Martina; White, Richard A.; Erschen, Sabine; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-08-06

    Streptomyces sp. strain Wb2n-11, isolated from native desert soil, exhibited broad-spectrum antagonism against plant pathogenic fungi, bacteria and nematodes. The 8.2 Mb draft genome reveals genes putatively responsible for its promising biocontrol activity and genes which enable the soil bacterium to directly interact beneficially with plants.

  9. Complete Genome Sequence of Micromonospora Strain L5, a Potential Plant-Growth-Regulating Actinomycete, Originally Isolated from Casuarina equisetifolia Root Nodules

    Energy Technology Data Exchange (ETDEWEB)

    Hirsch, A. M.; Alvarado, J.; Bruce, D.; Chertkov, O.; De Hoff, P. L.; Detter, J. C.; Fujishige, N. A.; Goodwin, L. A.; Han, J.; Han, S.; Ivanova, N.; Land, M. L.; Lum, M. R.; Milani-Nejad, N.; Nolan, M.; Pati, A.; Pitluck, S.; Tran, S. S.; Woyke, T.; Valdes, M.

    2013-08-29

    Micromonospora species live in diverse environments and exhibit a broad range of functions including antibiotic production, biocontrol, and ability to degrade complex polysaccharides. To learn more about these versatile actinomycetes, we sequenced the genome of strain L5, originally isolated from root nodules of an actinorhizal plant growing in Mexico.

  10. Ustilago maydis killer toxin as a new tool for the biocontrol of the wine spoilage yeast Brettanomyces bruxellensis.

    Science.gov (United States)

    Santos, Antonio; Navascués, Eva; Bravo, Enrique; Marquina, Domingo

    2011-01-31

    Brettanomyces bruxellensis is one of the most damaging species for wine quality, and tools for controlling its growth are limited. In this study, thirty-nine strains belonging to Saccharomyces cerevisiae and B. bruxellensis have been isolated from wineries, identified and then tested against a panel of thirty-nine killer yeasts. Here, for the first time, the killer activity of Ustilago maydis is proven to be effective against B. bruxellensis. Mixed cultures in winemaking conditions show that U. maydis CYC 1410 has the ability to inhibit B. bruxellensis, while S. cerevisiae is fully resistant to its killer activity, indicating that it could be used in wine fermentation to avoid the development of B. bruxellensis without undesirable effects on the fermentative yeast. The characterization of the dsRNAs isolated and purified from U. maydis CYC 1410 indicated that this strain produces a KP6-related toxin. Killer toxin extracts were active against B. bruxellensis at pH values between 3.0 and 4.5 and temperatures comprised between 15 °C and 25 °C, confirming their biocontrol activity in winemaking and wine aging conditions. Furthermore, small amounts (100 AU/ml) of killer toxin extracts from U. maydis significantly reduced the amount of 4-ethylphenol produced by B. bruxellensis, indicating that in addition to the growth inhibition observed for high killer toxin concentrations (ranging from 400 to 2000 AU/ml), small amounts of the toxin are able to reduce the production of volatile phenols responsible for the aroma defects in wines caused by B. bruxellensis.

  11. Augmentative Biocontrol in Natural Marine Habitats: Persistence, Spread and Non-Target Effects of the Sea Urchin Evechinus chloroticus

    OpenAIRE

    Javier Atalah; Hopkins, Grant A.; Forrest, Barrie M.

    2013-01-01

    Augmentative biocontrol aims to control established pest populations through enhancement of their indigenous enemies. To our knowledge, this approach has not been applied at an operational scale in natural marine habitats, in part because of the perceived risk of adverse non-target effects on native ecosystems. In this paper, we focus on the persistence, spread and non-target effects of the sea urchin Evechinus chloroticus when used as biocontrol agent to eradicate an invasive kelp from Fiord...

  12. Biocontrole de Listeria monocytogenes por Pediococcus acidilactici em couve minimamente processada Biocontrol of Listeria monocytogenes by Pediococcus acidilactici in fresh-cut kale

    OpenAIRE

    Wanessa Altimiras Costa; Maria Cristina Dantas Vanetti; Rolf Puschamann

    2009-01-01

    Este estudo avaliou um sistema de biocontrole para inibição de Listeria monocytogenes em couve minimamente processada, objetivando sua segurança durante estocagem sob refrigeração e em condições de abuso de temperatura. O potencial inibitório de bactérias láticas tolerantes ao sal e psicrotróficas contaminantes naturais da couve e Lactobacillus plantarum, Lactobacillus delbrueckii ATCC 9649 e Lactobacillus casei CCT 1465 foram avaliadas contra L. monocytogenes. O isolado de couve identificado...

  13. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7

    Science.gov (United States)

    Maldonado-González, M. Mercedes; Bakker, Peter A. H. M.; Prieto, Pilar; Mercado-Blanco, Jesús

    2015-01-01

    The effective management of Verticillium wilts (VW), diseases affecting many crops and caused by some species of the soil-borne fungus Verticillium, is problematic. The use of microbial antagonists to control these pathologies fits modern sustainable agriculture criteria. Pseudomonas fluorescens PICF7 is an endophytic bacterium isolated from olive roots with demonstrated ability to control VW of olive caused by the highly virulent, defoliating (D) pathotype of Verticillium dahliae Kleb. However, the study of the PICF7-V. dahliae-olive tripartite interaction poses difficulties because of the inherent characteristics of woody, long-living plants. To overcome these problems we explored the use of the model plant Arabidopsis thaliana. Results obtained in this study showed that: (i) olive D and non-defoliating V. dahliae pathotypes produce differential disease severity in A. thaliana plants; (ii) strain PICF7 is able to colonize and persist in the A. thaliana rhizosphere but is not endophytic in Arabidopsis; and (iii) strain PICF7 controls VW in Arabidopsis. Additionally, as previously observed in olive, neither swimming motility nor siderophore production by PICF7 are required for VW control in A. thaliana, whilst cysteine auxotrophy decreased the effectiveness of PICF7. Moreover, when applied to the roots PICF7 controlled Botrytis cinerea infection in the leaves of Arabidopsis, suggesting that this strain is able to induce systemic resistance. A. thaliana is therefore a suitable alternative to olive bioassays to unravel biocontrol traits involved in biological control of V. dahliae by P. fluorescens PICF7. PMID:25904904

  14. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7.

    Science.gov (United States)

    Maldonado-González, M Mercedes; Bakker, Peter A H M; Prieto, Pilar; Mercado-Blanco, Jesús

    2015-01-01

    The effective management of Verticillium wilts (VW), diseases affecting many crops and caused by some species of the soil-borne fungus Verticillium, is problematic. The use of microbial antagonists to control these pathologies fits modern sustainable agriculture criteria. Pseudomonas fluorescens PICF7 is an endophytic bacterium isolated from olive roots with demonstrated ability to control VW of olive caused by the highly virulent, defoliating (D) pathotype of Verticillium dahliae Kleb. However, the study of the PICF7-V. dahliae-olive tripartite interaction poses difficulties because of the inherent characteristics of woody, long-living plants. To overcome these problems we explored the use of the model plant Arabidopsis thaliana. Results obtained in this study showed that: (i) olive D and non-defoliating V. dahliae pathotypes produce differential disease severity in A. thaliana plants; (ii) strain PICF7 is able to colonize and persist in the A. thaliana rhizosphere but is not endophytic in Arabidopsis; and (iii) strain PICF7 controls VW in Arabidopsis. Additionally, as previously observed in olive, neither swimming motility nor siderophore production by PICF7 are required for VW control in A. thaliana, whilst cysteine auxotrophy decreased the effectiveness of PICF7. Moreover, when applied to the roots PICF7 controlled Botrytis cinerea infection in the leaves of Arabidopsis, suggesting that this strain is able to induce systemic resistance. A. thaliana is therefore a suitable alternative to olive bioassays to unravel biocontrol traits involved in biological control of V. dahliae by P. fluorescens PICF7. PMID:25904904

  15. Disruption of gene pqqA or pqqB reduces plant growth promotion activity and biocontrol of crown gall disease by Rahnella aquatilis HX2.

    Directory of Open Access Journals (Sweden)

    Lei Li

    Full Text Available Rahnella aquatilis strain HX2 has the ability to promote maize growth and suppress sunflower crown gall disease caused by Agrobacterium vitis, A. tumefaciens, and A. rhizogenes. Pyrroloquinoline quinone (PQQ, a cofactor of aldose and alcohol dehydrogenases, is required for the synthesis of an antibacterial substance, gluconic acid, by HX2. Mutants of HX2 unable to produce PQQ were obtained by in-frame deletion of either the pqqA or pqqB gene. In this study, we report the independent functions of pqqA and pqqB genes in relation to PQQ synthesis. Interestingly, both the pqqA and pqqB mutants of R. aquatilis eliminated the ability of strain HX2 to produce antibacterial substance, which in turn, reduced the effectiveness of the strain for biological control of sunflower crown gall disease. The mutation also resulted in decreased mineral phosphate solubilization by HX2, which reduced the efficacy of this strain as a biological fertilizer. These functions were restored by complementation with the wild-type pqq gene cluster. Additionally, the phenotypes of HX2 derivatives, including colony morphology, growth dynamic, and pH change of culture medium were impacted to different extents. Our findings suggested that pqqA and pqqB genes individually play important functions in PQQ biosynthesis and are required for antibacterial activity and phosphorous solubilization. These traits are essential for R. aquatilis efficacy as a biological control and plant growth promoting strain. This study enhances our fundamental understanding of the biosynthesis of an environmentally significant cofactor produced by a promising biocontrol and biological fertilizer strain.

  16. Combining UV-C treatment with biocontrol yeast to control postharvest decay of melon.

    Science.gov (United States)

    Huang, Ke; Zou, Yong; Luo, Jie; Liu, Yiqing

    2015-09-01

    Significant losses in harvested melon can be directly attributable to decay fungi. In the present study, the use of UV-C treatment combined with biocontrol yeast, Pichia cecembensis, was evaluated for their ability to control postharvest decay of melon fruits after they were artificially inoculated with Fusarium oxysporum and Alternaria alternata. Natural infection of fruit was also assessed. As a stand-alone treatment, UV-C or P. cecembensis significantly reduced Fusarium rot and Alternaria rot, and also the level of natural infection on melon fruit, relative to the untreated control. The combination of UV-C or P. cecembensis, however, provided a superior level of decay control on artificially inoculated and naturally infected fruit, compared to either treatment alone. None of the treatments impaired fruit quality. Integrating the use of UV-C treatment with biocontrol yeast has potential as an effective method to control postharvest decay of melon.

  17. Biocontrol Efficacy of Two Antagonistic Yeasts Against Postharvest Diseases in Peach Fruits During Storage Periods

    Institute of Scientific and Technical Information of China (English)

    LIN Li; TIAN Shi-ping; QIN Guo-zheng; XU Yong

    2003-01-01

    Two antagonistic yeasts, Thichosporon pullulans and Cryptococcus laurentii, were investigated for their biocontrol potential to blue mold rot and rhizopus rot on harvested peach fruits (Prunus persica L. Batsch, cv. Okubao), alone or in combination with a Iow dose of iprodione (50 μg mi-1 ). The results indicated that T. pullulans and C. laurentii were effective at reducing disease incidence and severity of blue mold rot and rhizopus rot in peach fruits. Biocontrol efficacy of C. laurentii and T. pullulans were significantly enhanced by combination with a Iow dose of iprodione (50 μg mi-1) against blue mold and rhizopus rot in peach fruits. T. pullulans and C. laurentii combined with a low dose of iprodione (50 μg mi-1 ) resulted in better disease control than either iprodione or the yeasts used alone. Dipping fruits in suspensions of antagonist cells showed the similar control effect as the treatment with iprodione (500 μg ml-1 ).

  18. Trichoderma spp.capable of growing at low temperatures with biocontrol potential

    Institute of Scientific and Technical Information of China (English)

    Szakacs G; Tavaszi A

    2004-01-01

    @@ Though there are successful commercial biocontrol products with Trichoderma spp. In many countries including US, Israel and Europe, their usefulness is limited in cold environments such as mid-and northern part of Europe, US and Canada, especially in the late fall, winter and early spring period.Trichoderma isolates capable of growing at low temperatures (5-10 ℃) and showing good antagonistic properties against plant pathogenic fungi may have therefore both scientific and commercial value.

  19. Enhanced biocontrol activity of Trichoderma through inactivation of a mitogen-activated protein kinase

    OpenAIRE

    Mendoza-Mendoza, Artemio; Pozo, María J.; Grzegorski, Darlene; Martínez, Pedro; García, Juan M.; Olmedo-Monfil, Vianey; Cortés, Carlos; Kenerley, Charles; Herrera-Estrella, Alfredo

    2003-01-01

    The production of lytic enzymes in Trichoderma is considered determinant in its parasitic response against fungal species. A mitogen-activated protein kinase encoding gene, tvk1, from Trichoderma virens was cloned, and its role during the mycoparasitism, conidiation, and biocontrol was examined in tvk1 null mutants. These mutants showed a clear increase in the level of the expression of mycoparasitism-related genes under simulated mycoparasitism and during direct confrontation with the plant ...

  20. Ecophysiological approaches to production and formulation of the biocontrol yeast Pichia anomala

    OpenAIRE

    Mokiou, Stella

    2004-01-01

    To produce commercial biocontrol agents (BCAs) successfully, it is important that cheap and economic substrates are used which support high numbers of good quality inoculum. Production of formulations conserving ecological competence and shelf-life should also be ensured. With this in mind, studies focusing on yeast ecophysiology were conducted to produce and formulate ecologically competent P. anomala cells for controlling spoilage of moist cereal grain. The liquid culture systems used were ...

  1. Ecology of Anti-Biofilm Agents II: Bacteriophage Exploitation and Biocontrol of Biofilm Bacteria

    OpenAIRE

    Abedon, Stephen T.

    2015-01-01

    Bacteriophages are the viruses of bacteria. In the guise of phage therapy they have been used for decades to successfully treat what are probable biofilm-containing chronic bacterial infections. More recently, phage treatment or biocontrol of biofilm bacteria has been brought back to the laboratory for more rigorous assessment as well as towards the use of phages to combat environmental biofilms, ones other than those directly associated with bacterial infections. Considered in a companion ar...

  2. Enhancement of biocontrol efficacy against Botrytis cinerea through the manipulation of nitrogen fertilization of tomato plants

    OpenAIRE

    Abro, Manzoor Ali; Lecompte, François; Bardin, Marc; Duffaud, Magali

    2013-01-01

    Although nitrogen fertilization is known to affect plant susceptibility to certain pathogens, little is known on its possible effect on the efficacy of biological control. In the present study we examined the effect of five levels of NO3- nutrition on the efficacy of two biocontrol agents (Trichoderma harzianum and Microdochium dimerum) to protect pruning wounds of tomato against Botrytis cinerea. Plants were grown for two months in a greenhouse with a soil-less drip-irrigationsystem. Differe...

  3. What Magnitude Are Observed Non-Target Impacts from Weed Biocontrol?

    OpenAIRE

    David Maxwell Suckling; René François Henri Sforza

    2014-01-01

    A systematic review focused by plant on non-target impacts from agents deliberately introduced for the biological control of weeds found significant non-target impacts to be rare. The magnitude of direct impact of 43 biocontrol agents on 140 non-target plants was retrospectively categorized using a risk management framework for ecological impacts of invasive species (minimal, minor, moderate, major, massive). The vast majority of agents introduced for classical biological control of weeds (>9...

  4. Statistical media optimization for the biomass production of postharvest biocontrol yeast Rhodosporidium paludigenum.

    Science.gov (United States)

    Wang, Peng; Liu, Xia; Wang, Yifei; Ruan, Hui; Zheng, XiaoDong

    2011-01-01

    A cane molasses-based medium for the biomass production of biocontrol agent Rhodosporidium paludigenum was statistically optimized. Molasses concentration (after pretreatment), yeast extract, and initial pH were identified by the Plackett-Burman design to show significant influence on the biomass production. The three factors were further optimized by central composite design and response-surface methodology. The statistical analysis indicated the optimum values of the variables were 89.98 g/L for cane molasses, 2.35 g/L for yeast extract and an initial pH of 8.48. The biomass yield at the optimal culture achieved 15.89 g/L in flask fermentation, which was 2.1 times higher than that at the initial NYDB medium. In a 10-L fermenter, 18.97 g/L of biomass was obtained after 36 hr of cultivation. Moreover, the biocontrol efficacy of the yeast was investigated after culture optimization. The results showed the yeast harvested in the optimal medium maintained its initial biocontrol properties by reducing the percentage of decayed apples to below 20%.

  5. Statistical media optimization for the biomass production of postharvest biocontrol yeast Rhodosporidium paludigenum.

    Science.gov (United States)

    Wang, Peng; Liu, Xia; Wang, Yifei; Ruan, Hui; Zheng, XiaoDong

    2011-01-01

    A cane molasses-based medium for the biomass production of biocontrol agent Rhodosporidium paludigenum was statistically optimized. Molasses concentration (after pretreatment), yeast extract, and initial pH were identified by the Plackett-Burman design to show significant influence on the biomass production. The three factors were further optimized by central composite design and response-surface methodology. The statistical analysis indicated the optimum values of the variables were 89.98 g/L for cane molasses, 2.35 g/L for yeast extract and an initial pH of 8.48. The biomass yield at the optimal culture achieved 15.89 g/L in flask fermentation, which was 2.1 times higher than that at the initial NYDB medium. In a 10-L fermenter, 18.97 g/L of biomass was obtained after 36 hr of cultivation. Moreover, the biocontrol efficacy of the yeast was investigated after culture optimization. The results showed the yeast harvested in the optimal medium maintained its initial biocontrol properties by reducing the percentage of decayed apples to below 20%. PMID:21967338

  6. Striga Biocontrol on a Toothpick: A Readily Deployable and Inexpensive Method for Smallholder Farmers

    Directory of Open Access Journals (Sweden)

    David Chandler Sands

    2016-08-01

    Full Text Available Striga hermonthica (witchweed is a parasitic weed that attacks and significantly reduces the yields of maize, sorghum, millet and sugarcane throughout sub-Saharan Africa. Low cost management methods such as hand weeding, short crop rotations, trap cropping, or conventional biocontrol have not been effective. Likewise, Striga-tolerant or herbicide-resistant maize cultivars are higher yielding, but are often beyond the economic means of sustenance farmers. The fungal pathogen, Fusarium oxysporum f.sp. strigae, has been the object of numerous studies to develop Striga biocontrol. Under experimental conditions this pathogen can reduce the incidence of Striga infestation but field use is not extensive, perhaps because it has not been sufficiently effective in restoring crop yield and reducing the soil Striga seed bank. Here we brought together Kenyan and US crop scientists with smallholder farmers to develop and validate an effective biocontrol strategy for management of Striga on smallholder farms. Key components of this research project were the following: 1. Development of a two-step method of fungal delivery, including laboratory coating of primary inoculum on toothpicks, followed by on-farm production of secondary field inoculum in boiled rice enabling delivery of vigorous, fresh inoculum directly to the seedbed; 2. Training of smallholder farmers (85% women, to produce the biocontrol agent and incorporate it into their maize plantings in Striga-infested soils and collect agronomic data. The field tests expanded from 30 smallholder farmers to a two-season, 500-farmer plot trial including paired plus and minus biocontrol plots with fertilizer and hybrid seed in both plots and; 3. Concerted selection of variants of the pathogen identified for enhanced virulence, as has been demonstrated in other host parasite systems were employed here on Striga via pathogen excretion of the amino acids L-leucine and L-tyrosine that are toxic to Striga but

  7. Striga Biocontrol on a Toothpick: A Readily Deployable and Inexpensive Method for Smallholder Farmers

    Science.gov (United States)

    Nzioki, Henry S.; Oyosi, Florence; Morris, Cindy E.; Kaya, Eylul; Pilgeram, Alice L.; Baker, Claire S.; Sands, David C.

    2016-01-01

    Striga hermonthica (witchweed) is a parasitic weed that attacks and significantly reduces the yields of maize, sorghum, millet, and sugarcane throughout sub-Saharan Africa. Low cost management methods such as hand weeding, short crop rotations, trap cropping, or conventional biocontrol have not been effective. Likewise, Striga-tolerant or herbicide-resistant maize cultivars are higher yielding, but are often beyond the economic means of sustenance farmers. The fungal pathogen, Fusarium oxysporum f.sp. strigae, has been the object of numerous studies to develop Striga biocontrol. Under experimental conditions this pathogen can reduce the incidence of Striga infestation but field use is not extensive, perhaps because it has not been sufficiently effective in restoring crop yield and reducing the soil Striga seed bank. Here we brought together Kenyan and US crop scientists with smallholder farmers to develop and validate an effective biocontrol strategy for management of Striga on smallholder farms. Key components of this research project were the following: (1) Development of a two-step method of fungal delivery, including laboratory coating of primary inoculum on toothpicks, followed by on-farm production of secondary field inoculum in boiled rice enabling delivery of vigorous, fresh inoculum directly to the seedbed; (2) Training of smallholder farmers (85% women), to produce the biocontrol agent and incorporate it into their maize plantings in Striga-infested soils and collect agronomic data. The field tests expanded from 30 smallholder farmers to a two-season, 500-farmer plot trial including paired plus and minus biocontrol plots with fertilizer and hybrid seed in both plots and; (3) Concerted selection of variants of the pathogen identified for enhanced virulence, as has been demonstrated in other host parasite systems were employed here on Striga via pathogen excretion of the amino acids L-leucine and L-tyrosine that are toxic to Striga but innocuous to maize

  8. Striga Biocontrol on a Toothpick: A Readily Deployable and Inexpensive Method for Smallholder Farmers.

    Science.gov (United States)

    Nzioki, Henry S; Oyosi, Florence; Morris, Cindy E; Kaya, Eylul; Pilgeram, Alice L; Baker, Claire S; Sands, David C

    2016-01-01

    Striga hermonthica (witchweed) is a parasitic weed that attacks and significantly reduces the yields of maize, sorghum, millet, and sugarcane throughout sub-Saharan Africa. Low cost management methods such as hand weeding, short crop rotations, trap cropping, or conventional biocontrol have not been effective. Likewise, Striga-tolerant or herbicide-resistant maize cultivars are higher yielding, but are often beyond the economic means of sustenance farmers. The fungal pathogen, Fusarium oxysporum f.sp. strigae, has been the object of numerous studies to develop Striga biocontrol. Under experimental conditions this pathogen can reduce the incidence of Striga infestation but field use is not extensive, perhaps because it has not been sufficiently effective in restoring crop yield and reducing the soil Striga seed bank. Here we brought together Kenyan and US crop scientists with smallholder farmers to develop and validate an effective biocontrol strategy for management of Striga on smallholder farms. Key components of this research project were the following: (1) Development of a two-step method of fungal delivery, including laboratory coating of primary inoculum on toothpicks, followed by on-farm production of secondary field inoculum in boiled rice enabling delivery of vigorous, fresh inoculum directly to the seedbed; (2) Training of smallholder farmers (85% women), to produce the biocontrol agent and incorporate it into their maize plantings in Striga-infested soils and collect agronomic data. The field tests expanded from 30 smallholder farmers to a two-season, 500-farmer plot trial including paired plus and minus biocontrol plots with fertilizer and hybrid seed in both plots and; (3) Concerted selection of variants of the pathogen identified for enhanced virulence, as has been demonstrated in other host parasite systems were employed here on Striga via pathogen excretion of the amino acids L-leucine and L-tyrosine that are toxic to Striga but innocuous to maize

  9. Identification of Biocontrol Bacteria against Soybean Root Rot with Biolog Automatic Microbiology Analysis System%拮抗大豆根腐病细菌的Biolog鉴定

    Institute of Scientific and Technical Information of China (English)

    许艳丽; 刘海龙; 李春杰; 潘凤娟; 李淑娴; 刘新晶

    2012-01-01

    In order to identify the systematic position of taxonomy of two biocontrol bacteria against soybean root rot. Traditional morphological identification and BIOLOG automatic microbiology analysis system were used to identify strain B021a and B04b. The results showed that similarity value of strain B021a with Vibrio tubiashii was 0. 634, possibility to 86% and genetic distance to 4.00,and similarity value of strain B04b with Pasteurella trehalosi was 0. 610,probability to 75% and genetic distance to 2. 77. Strain B021a was identified as Vibrio tubiashii and strain B04b as Pasteurella trehalosi by colony morphological propertie and BIOLOC analysis system.%为明确2株生防细菌的分类地位,采用传统形态学方法结合Biolog微生物自动分析系统,鉴定了大豆根腐病的2株生防细菌.结果表明,菌株B021a与塔式弧菌相似度值为0.634,可能性是86%,遗传距离为4.00.菌株B04b与海藻巴斯德菌相似度值为0.610,可能性是75%,遗传距离为2.77.综合形态学和Biolog鉴定结果,认为菌株B021a是塔式弧菌,菌株B04b是海藻巴斯德菌.

  10. Peptaibol, secondary-metabolite, and hydrophobin pattern of commercial biocontrol agents formulated with species of the Trichoderma harzianum complex.

    Science.gov (United States)

    Degenkolb, Thomas; Fog Nielsen, Kristian; Dieckmann, Ralf; Branco-Rocha, Fabiano; Chaverri, Priscila; Samuels, Gary J; Thrane, Ulf; von Döhren, Hans; Vilcinskas, Andreas; Brückner, Hans

    2015-04-01

    The production of bioactive polypeptides (peptaibiotics) in vivo is a sophisticated adaptation strategy of both mycoparasitic and saprotrophic Trichoderma species for colonizing and defending their natural habitats. This feature is of major practical importance, as the detection of peptaibiotics in plant-protective Trichoderma species, which are successfully used against economically relevant bacterial and fungal plant pathogens, certainly contributes to a better understanding of these complex antagonistic interactions. We analyzed five commercial biocontrol agents (BCAs), namely Canna(®) , Trichosan(®) , Vitalin(®) , Promot(®) WP, and TrichoMax(®) , formulated with recently described species of the Trichoderma harzianum complex, viz. T. afroharzianum, T. simmonsii, and T. guizhouense. By using the well-established, HPLC/MS-based peptaibiomics approach, it could unequivocally be demonstrated that all of these formulations contained new and recurrent peptaibols, i.e., peptaibiotics carrying an acetylated N-terminus, the C-terminus of which is reduced to a 1,2-amino alcohol. Their chain lengths, including the amino alcohol, were 11, 14, and 18 residues, respectively. Peptaibols were also to be the dominating secondary metabolites in plate cultures of the four strains obtained from four of the Trichoderma- based BCAs, contributing 95% of the UHPLC-UV/VIS peak areas and 99% of the total ion count MS peak area from solid media. Furthermore, species-specific hydrophobins, as well as non-peptaibiotic secondary metabolites, were detected, the latter being known for their antifungal, siderophore, or plant-growth-promoting activities. Notably, none of the isolates produced low-molecular weight mycotoxins.

  11. Functional analysis of the class II hydrophobin gene HFB2-6 from the biocontrol agent Trichoderma asperellum ACCC30536.

    Science.gov (United States)

    Huang, Ying; Mijiti, Gulijimila; Wang, Zhiying; Yu, Wenjing; Fan, Haijuan; Zhang, Rongshu; Liu, Zhihua

    2015-02-01

    A class II hydrophobin gene, HFB2-6, was cloned from Trichoderma asperellum ACCC30536 and its biocontrol function was studied. According to our previous transcriptome data, six of the eight class II hydrophobin genes were obviously differential expression in four inducing conditions, especially the gene HFB2-6. Moreover, HFB2-6 proven to be differentially transcribed under eight different treatments. HFB2-6 transcripts were up-regulated under 1% Alternaria alternata cell wall and 5% A. alternata fermentation liquid treatments, and by nutritional stress conditions, suggesting that HFB2-6 plays roles in interactions with both biotic and abiotic environmental conditions. HFB2-6 expression was down-regulated under 1% poplar leaf powder culture conditions, but its expression was up-regulated under 1% poplar root powder, indicating that HFB2-6 has a function in root colonization. Furthermore, the recombinant hydrophobin rHFB2-6 was successfully expressed in Escherichia coli BL21-HFB2-6 and purified from the recombinant strain. Genes related to both the jasmonic acid and salicylic acid signal transduction pathways were up-regulated by interaction with renatured rHFB2-6. The ORCA3 (octadecanoid-derivative responsive Catharanthus AP2-domain) gene of the poplar jasmonic acid signal transduction pathway showed a peak expression of 4.48 times at 2 h, and the peak expression of PR1 (pathogenesis-related protein gene) in the salicylic acid signal transduction pathway was 4.58 times at 72 h. Two genes, MP (monopteros) and GH3.17 (auxin original response gene), in the auxin signal transduction pathway were also up-regulated after induction with rHFB2-6, indicating that rHFB2-6 can promote poplar growth and confer broad-spectrum resistance to pathogens.

  12. Peptaibol, secondary-metabolite, and hydrophobin pattern of commercial biocontrol agents formulated with species of the Trichoderma harzianum complex.

    Science.gov (United States)

    Degenkolb, Thomas; Fog Nielsen, Kristian; Dieckmann, Ralf; Branco-Rocha, Fabiano; Chaverri, Priscila; Samuels, Gary J; Thrane, Ulf; von Döhren, Hans; Vilcinskas, Andreas; Brückner, Hans

    2015-04-01

    The production of bioactive polypeptides (peptaibiotics) in vivo is a sophisticated adaptation strategy of both mycoparasitic and saprotrophic Trichoderma species for colonizing and defending their natural habitats. This feature is of major practical importance, as the detection of peptaibiotics in plant-protective Trichoderma species, which are successfully used against economically relevant bacterial and fungal plant pathogens, certainly contributes to a better understanding of these complex antagonistic interactions. We analyzed five commercial biocontrol agents (BCAs), namely Canna(®) , Trichosan(®) , Vitalin(®) , Promot(®) WP, and TrichoMax(®) , formulated with recently described species of the Trichoderma harzianum complex, viz. T. afroharzianum, T. simmonsii, and T. guizhouense. By using the well-established, HPLC/MS-based peptaibiomics approach, it could unequivocally be demonstrated that all of these formulations contained new and recurrent peptaibols, i.e., peptaibiotics carrying an acetylated N-terminus, the C-terminus of which is reduced to a 1,2-amino alcohol. Their chain lengths, including the amino alcohol, were 11, 14, and 18 residues, respectively. Peptaibols were also to be the dominating secondary metabolites in plate cultures of the four strains obtained from four of the Trichoderma- based BCAs, contributing 95% of the UHPLC-UV/VIS peak areas and 99% of the total ion count MS peak area from solid media. Furthermore, species-specific hydrophobins, as well as non-peptaibiotic secondary metabolites, were detected, the latter being known for their antifungal, siderophore, or plant-growth-promoting activities. Notably, none of the isolates produced low-molecular weight mycotoxins. PMID:25879509

  13. Chitinolytic Bacteria Isolated from Chili Rhizosphere: Chitinase Characterization and Its Application as Biocontrol for Whitefly (Bemisia tabaci Genn.

    Directory of Open Access Journals (Sweden)

    Nisa R. Mubarik

    2010-01-01

    Full Text Available Problem statement: Chitin, a common constituent of insect exoskeleton, could be hydrolyzed by chitinase. The research was conducted to screen chitinolytic rhizobacteria isolated from rhizosphere of chilli pepper and to determine their chitinase activity in degrading chitin of whitefly, Bemisia tabaci Genn. (Hemiptera: Aleyrodidae. Whitefly is recognized as an important pest on many crops including chilli pepper. Approach: Screening and molecular identification based on 16S rRNA sequence of chitinolytic isolates, chitinase productions, measurement of chitinase activity, characterization of chitinase and effect of the chitinase treatment on whitefly were studied. Results: A total of 25 isolates of rhizobacteria formed a clear zone on solid chitin media. Two isolates, i.e., I.5 and I.21 isolates had the highest chitinolytic index. Based on sequence of 16S rRNA gene, the isolates of I.5 and I.21 were identified as Bacillus sp. and Bacillus cereus, respectively. The highest chitinolytic index and specific activity of I.5 isolate was 0.94 and 0.11 U mg-1 proteins, respectively. Maximum production of I.5 chitinase was occured after 36 h cultivation at 30°C and pH 7.0. The highest chitinolytic index and specific activity of I.21isolate was 0.75 and 0.114 U mg-1 proteins, respectively. Maximum production of I.21 chitinase was occured after 36 h cultivation at 55°C and pH 7.0. Cell culture and crude enzyme of the isolates were tested on chitin of B. tabaci and the effect was observed using a microscope and sterile water was used as a negative control. Hydrolytic observation showed that crude enzyme of I.21 isolate could degrade chitin of B. tabaci exoskeleton and the activity was better than that of I.5 isolate. Conclusion: Chitinase produced by Bacillus cereus I.21 strain has potential application as biocontrol agents for B. tabaci.

  14. Colonization process of olive tissues by Verticillium dahliae and its in planta interaction with the biocontrol root endophyte Pseudomonas fluorescens PICF7

    Science.gov (United States)

    Prieto, Pilar; Navarro‐Raya, Carmen; Valverde‐Corredor, Antonio; Amyotte, Stefan G.; Dobinson, Katherine F.; Mercado‐Blanco, Jesús

    2009-01-01

    Summary The colonization process of Olea europaea by the defoliating pathotype of Verticillium dahliae, and the in planta interaction with the endophytic, biocontrol strain Pseudomonas fluorescens PICF7 were determined. Differential fluorescent protein tagging was used for the simultaneous visualization of P. fluorescens PICF7 and V. dahliae in olive tissues. Olive plants were bacterized with PICF7 and then transferred to V. dahliae‐infested soil. Monitoring olive colonization events by V. dahliae and its interaction with PICF7 was conducted using a non‐gnotobiotic system, confocal laser scanner microscopy and tissue vibratoming sections. A yellow fluorescently tagged V. dahliae derivative (VDAT‐36I) was obtained by Agrobacterium tumefaciens‐mediated transformation. Isolate VDAT‐36I quickly colonized olive root surface, successfully invaded root cortex and vascular tissues via macro‐ and micro‐breakages, and progressed to the aerial parts of the plant through xylem vessel cells. Strain PICF7 used root hairs as preferred penetration site, and once established on/in root tissues, hindered pathogen colonization. For the first time using this approach, the entire colonization process of a woody plant by V. dahliae is reported. Early and localized root surface and root endophytic colonization by P. fluorescens PICF7 is needed to impair full progress of verticillium wilt epidemics in olive. PMID:21255281

  15. Zoospore homing and infection events: effects of the biocontrol bacterium Burkholderia cepacia AMMDR1 on two oomycete pathogens of pea (Pisum sativum L.).

    Science.gov (United States)

    Heungens, K; Parke, J L

    2000-12-01

    Burkholderia cepacia AMMDR1 is a biocontrol agent that protects pea and sweet corn seeds from Pythium damping-off in field experiments. The goal of this work was to understand the effect of B. cepacia AMMDR1 on Pythium aphanidermatum and Aphanomyces euteiches zoospore homing events and on infection of pea seeds or roots. In vitro, B. cepacia AMMDR1 caused zoospore lysis, prevented cyst germination, and inhibited germ tube growth of both oomycetes. B. cepacia AMMDR1 also reduced the attractiveness of seed exudates to Pythium zoospores to nondetectable levels. However, when present at high levels on seeds, B. cepacia AMMDR1 had little net effect on zoospore attraction, probably because it also enhanced seed exudation. Seed-applied B. cepacia AMMDR1 dramatically reduced the incidence of infection by Pythium zoospores in situ compared with an antibiosis-deficient Tn5 mutant strain. This mutant strain also decreased Pythium infection incidence to some extent, but only when the pathogen inoculum potential was low. B. cepacia AMMDR1 did not affect attraction of Aphanomyces zoospores or Aphanomyces root rot incidence. These results suggest that B. cepacia AMMDR1 controls P. aphanidermatum largely through antibiosis, but competition for zoospore-attracting compounds can contribute to the effect. Differences in suppression of Aphanomyces and Pythium are discussed in relation to differences in the ecology of the two pathogens. PMID:11097889

  16. Biocontrol of Rhizoctonia solani damping-off and promotion of tomato plant growth by endophytic actinomycetes isolated from native plants of Algerian Sahara.

    Science.gov (United States)

    Goudjal, Yacine; Toumatia, Omrane; Yekkour, Amine; Sabaou, Nasserdine; Mathieu, Florence; Zitouni, Abdelghani

    2014-01-20

    Thirty-four endophytic actinomycetes were isolated from the roots of native plants of the Algerian Sahara. Morphological and chemical studies showed that twenty-nine isolates belonged to the Streptomyces genus and five were non-Streptomyces. All isolates were screened for their in vitro antifungal activity against Rhizoctonia solani. The six that had the greatest pathogen inhibitory capacities were subsequently tested for their in vivo biocontrol potential on R. solani damping-off in sterilized and non-sterilized soils, and for their plant-growth promoting activities on tomato seedlings. In both soils, coating tomato seeds with antagonistic isolates significantly reduced (Pdamping-off of tomato seedlings. Among the isolates tested, the strains CA-2 and AA-2 exhibited the same disease incidence reduction as thioperoxydicarbonic diamide, tetramethylthiram (TMTD) and no significant differences (Pseed-treated seedlings compared to the control. The taxonomic position based on 16S rDNA sequence analysis and phylogenetic studies indicated that the strains CA-2 and AA-2 were related to Streptomyces mutabilis NBRC 12800(T) (100% of similarity) and Streptomyces cyaneofuscatus JCM 4364(T) (100% of similarity), respectively. PMID:23920229

  17. Identification of potential marker genes for Trichoderma harzianum strains with high antagonistic potential against Rhizoctonia solani by a rapid subtraction hybridization approach.

    Science.gov (United States)

    Scherm, Barbara; Schmoll, Monika; Balmas, Virgilio; Kubicek, Christian P; Migheli, Quirico

    2009-02-01

    A rapid subtraction hybridization approach was used to isolate genes differentially expressed during mycelial contact between Trichoderma harzianum (Hypocrea lixii) and Rhizoctonia solani, and could serve as marker genes for selection of superior biocontrol strains. Putatively positive clones were evaluated by transcription analysis during mycelial contact with R. solani versus growth on glucose, and for their differential transcription between two strains with either strong or poor biocontrol capability before, at, and after contact with R. solani. Besides four clones, which had similarity to putative but as yet uncharacterized proteins, they comprised ribosomal proteins, proteins involved in transcriptional switch and regulation, amino acid and energy catabolism, multidrug resistance, and degradation of proteins and glucans. Transcription of three clones was evaluated in five T. harzianum strains under confrontation conditions with R. solani. Two clones-acetyl-xylane esterase AXE1 and endoglucanase Cel61b-showed significant upregulation during in vivo confrontation of a T. harzianum strain that successively demonstrated a very high antagonistic capability towards R. solani, while expression was progressively lower in a series of T. harzianum strains with intermediate to poor antagonistic activity. These clones are promising candidates for use as markers in the screening of improved T. harzianum biocontrol strains.

  18. Quantification of Pseudomonas fluorescens strains F113, CHA0 and Pf153 in the rhizosphere of maize by strain-specific real-time PCR unaffected by the variability of DNA extraction efficiency.

    Science.gov (United States)

    Von Felten, Andreas; Défago, Geneviève; Maurhofer, Monika

    2010-05-01

    Pseudomonas fluorescens strains F113 and CHA0 are well-known plant growth-promoting rhizobacteria (PGPR) often used as model strains in biocontrol experiments. To monitor their persistence in large scale field experiments, culture-independent methods are needed. In this study, a strain-specific real-time PCR quantification tool was developed based on sequence-characterized amplified regions (SCAR) for P. fluorescens strains F113, CHA0 and Pf153. Differences in DNA extraction efficiencies from rhizosphere samples were circumvented using plasmid APA9 as internal standard to normalize C(T) values after real-time amplification. The detection limits of the real-time PCR assays for all three strains were approximately 10 cells for genomic DNA and 10(4)cells/g rhizosphere for maize samples grown in different natural soils. Population sizes of the three strains in the rhizosphere of maize measured by the new real-time PCR approaches were similar to those measured by most probable number (MPN)-PCR. A persistence study of the three strains indicated that the strains persisted differently over a period of 5weeks. In conclusion the newly developed real-time PCR approach is a fast and resource efficient method for monitoring individual biocontrol strains in natural soil, which makes it an apt quantification tool for future large-scale field experiments. PMID:20153383

  19. Deciphering the conserved genetic loci implicated in plant disease control through comparative genomics of Bacillus amyloliquefaciens subsp. plantarum strains

    Directory of Open Access Journals (Sweden)

    Mohammad J Hossain

    2015-08-01

    Full Text Available To understand the growth-promoting and disease-inhibiting activities of plant growth-promoting rhizobacteria (PGPR strains, the genomes of 12 Bacillus subtilis group strains with PGPR activity were sequenced and analyzed. These B. subtilis strains exhibited high genomic diversity, whereas the genomes of B. amyloliquefaciens strains (a member of the B. subtilis group are highly conserved. A pairwise BLASTp matrix revealed that gene family similarity among Bacillus genomes ranges from 32- 90%, with 2,839 genes within the core genome of B. amyloliquefaciens subsp. plantarum. Comparative genomic analyses of B. amyloliquefaciens strains identified genes that are linked with biological control and colonization of roots and/or leaves, including 73 genes uniquely associated with subsp. plantarum strains that have predicted functions related to signaling, transportation, secondary metabolite production, and carbon source utilization. Although B. amyloliquefaciens subsp. plantarum strains contain gene clusters that encode many different secondary metabolites, only polyketide biosynthetic clusters that encode difficidin and macrolactin are conserved within this subspecies. To evaluate their role in plant pathogen biocontrol, genes involved in secondary metabolite biosynthesis were deleted in B. amyloliquefaciens subsp. plantarum strain, revealing that difficidin expression is critical in reducing the severity of disease, caused by Xanthomonas axonopodis pv. vesicatoria in tomato plants. This study defines genomic features of PGPR strains and links them with biocontrol activity and with host colonization.

  20. Bio-control potential of Cladosporium sp. (MCPL-461), against a noxious weed Parthenium hysterophorus L.

    Science.gov (United States)

    Kumar, Anuj; Verma, V C; Gond, S K; Kumar, V; Kharwar, R N

    2009-03-01

    The phenological survey of Parthenium hysterophorus L., in and around the campus of Banaras Hindu University (BHU) was done for about two years (2004-06). During Nov 2004, a few Parthenium plants were found diseased, and symptoms were restricted to the flowers, buds, and inflorescences. The disease causes sterility and reduces seed viability, which was observed with seed germination test from infected and healthy plants. The fungal pathogen was isolated and identified as Cladosporium sp. (MCPL-461). The severity of pathogen to the reproductive organs led to serious damages of the Parthenium plants. Thus in vitro and in vivo experiments were conducted to determine the bio-control potential of Cladosporium sp. (MCPL 461) against Parthenium weed. A combinatorial effort of Cladosporium sp. (MCPL 461) bio-control potential was evaluated with different culture media, incubation periods and spores strength. Spore suspension of 10(5) to 10(12) spores ml(-1) were used to spray on healthy Parthenium plants, and it was found that severe infection symptoms were appeared at 10(10) to 10(12) spores ml(-1) suspension. LD50 was found at 10(7) spores ml(-1). To enhance the myco-herbicide activity 3% sucrose was added to the spore suspension, which further resolute the bio-control efficacy of the isolates. Only 20-30% seeds of infected plants could germinate. However the safety of non-targeted and wild plants was also tested with Lantana camera, Chromolaena odorata and found that suspension up to 10(12) spores ml(-1) were not sufficient for disease outbreak in them. PMID:20121037

  1. Bio-control potential of Cladosporium sp. (MCPL-461), against a noxious weed Parthenium hysterophorus L.

    Science.gov (United States)

    Kumar, Anuj; Verma, V C; Gond, S K; Kumar, V; Kharwar, R N

    2009-03-01

    The phenological survey of Parthenium hysterophorus L., in and around the campus of Banaras Hindu University (BHU) was done for about two years (2004-06). During Nov 2004, a few Parthenium plants were found diseased, and symptoms were restricted to the flowers, buds, and inflorescences. The disease causes sterility and reduces seed viability, which was observed with seed germination test from infected and healthy plants. The fungal pathogen was isolated and identified as Cladosporium sp. (MCPL-461). The severity of pathogen to the reproductive organs led to serious damages of the Parthenium plants. Thus in vitro and in vivo experiments were conducted to determine the bio-control potential of Cladosporium sp. (MCPL 461) against Parthenium weed. A combinatorial effort of Cladosporium sp. (MCPL 461) bio-control potential was evaluated with different culture media, incubation periods and spores strength. Spore suspension of 10(5) to 10(12) spores ml(-1) were used to spray on healthy Parthenium plants, and it was found that severe infection symptoms were appeared at 10(10) to 10(12) spores ml(-1) suspension. LD50 was found at 10(7) spores ml(-1). To enhance the myco-herbicide activity 3% sucrose was added to the spore suspension, which further resolute the bio-control efficacy of the isolates. Only 20-30% seeds of infected plants could germinate. However the safety of non-targeted and wild plants was also tested with Lantana camera, Chromolaena odorata and found that suspension up to 10(12) spores ml(-1) were not sufficient for disease outbreak in them.

  2. [Fungal bio-control agents against ixodid tick--a review].

    Science.gov (United States)

    Sun, Ming; Wang, Xiaoyan; Luo, Jianxun

    2012-08-01

    Entomopathogenic fungi as potential agents for bio-control have been widely applied in the control of insect pests in agriculture. However, the application remains in laboratory scale for the control of ectoparasites. Owing to the need to combat the short lasting period of chemical acaricides and reduction of pollution, it is urgent to develop sufficient, stable and safe measures for tick control. We reviewed the primary scientific achievements in utilization of environmental microbes for controlling of ticks. Studies conducted in this field may benefit to sustainable development, environmental protection, maintaining ecological balance and production of green products.

  3. Involvement of Nitrate Reductase and Pyoverdine in Competitiveness of Pseudomonas fluorescens Strain C7R12 in Soil

    OpenAIRE

    Mirleau, Pascal; Philippot, Laurent; Corberand, Thérèse; Lemanceau, Philippe

    2001-01-01

    Involvement of nitrate reductase and pyoverdine in the competitiveness of the biocontrol strain Pseudomonas fluorescens C7R12 was determined, under gnotobiotic conditions, in two soil compartments (bulk and rhizosphere soil), with the soil being kept at two different values of matric potential (−1 and −10 kPa). Three mutants affected in the synthesis of either the nitrate reductase (Nar−), the pyoverdine (Pvd−), or both (Nar− Pvd−) were used. The Nar− and Nar− Pvd− mutants were obtained by si...

  4. Non-target trials with Pseudomonas fluorescens strain CL145A, a lethal control agent of dreissenid mussels (Bivalvia: Dreissenidae)

    OpenAIRE

    Molloy, Daniel P.; Denise A. Mayer; Michael J. Gaylo; Burlakova, Lyubov E.; Karatayev, Alexander Y.; Kathleen T. Presti; Paul M. Sawyko; John T. Morse; Eric A. Paul

    2013-01-01

    In an effort to develop an efficacious and environmentally safe method for managing zebra mussels (Dreissena polymorpha) and quaggamussels (Dreissena rostriformis bugensis), we initiated a research project investigating the potential use of bacteria and their naturalmetabolic products as biocontrol agents. This project resulted in the discovery of an environmental isolate lethal to dreissenid mussels,Pseudomonas fluorescens strain CL145A (Pf-CL145A). In previous published reports we have demo...

  5. Induction of Drought Tolerance in Cucumber Plants by a Consortium of Three Plant Growth-Promoting Rhizobacterium Strains

    OpenAIRE

    Chun-Juan Wang; Wei Yang; Chao Wang; Chun Gu; Dong-Dong Niu; Hong-Xia Liu; Yun-Peng Wang; Jian-Hua Guo

    2012-01-01

    Our previous work showed that a consortium of three plant growth-promoting rhizobacterium (PGPR) strains (Bacillus cereus AR156, Bacillus subtilis SM21, and Serratia sp. XY21), termed as BBS for short, was a promising biocontrol agent. The present study investigated its effect on drought tolerance in cucumber plants. After withholding watering for 13 days, BBS-treated cucumber plants had much darker green leaves and substantially lighter wilt symptoms than control plants. Compared to the cont...

  6. Biocontrol activity of Paenibacillus polymyxa AC-1 against Pseudomonas syringae and its interaction with Arabidopsis thaliana.

    Science.gov (United States)

    Hong, Chi Eun; Kwon, Suk Yoon; Park, Jeong Mee

    2016-04-01

    Paenibacillus polymyxa AC-1 (AC-1) is a plant growth-promoting rhizobacterium (PGPR) that has been used as a soil inoculant for biocontrol of plant pathogenic fungi and to promote plant growth. In this study, we examine the effects of AC-1 on the bacterial phytopathogen Pseudomonas syringae and internal colonization of AC-1 by counting bacterial populations that colonize plants. AC-1 inhibited the growth of both P. syringae pv. tomato DC3000 (Pst) and P. syringae pv. tabaci (Pta) in a concentration-dependent manner in in vitro assays. Upon treatment of AC-1 dropping at root tip of axenically grown Arabidopsis, we found that most of the AC-1 was detected in interior of leaves of Arabidiopsis plants rather than roots after 5 days post infection, indicating systemic spreading of AC-1 occur. We examined further AC-1 colonization patterns in Arabidopsis mutants deficient in phytohormone signaling pathways. These results indicated that abscisic acid (ABA) and jasmonic acid (JA) signaling pathways positively and negatively contributed, respectively, to AC-1 colonization of leaves, whereas epiphytic accumulation of AC-1 around root tissues was not affected. This study shows that AC-1 is an effective biocontrol agent to suppress P. syringae growth, possibly owing to its colonization patterns as a leaf-inhabiting endophyte. The results showed in this work will help to expand our understanding of the mode of action of AC-1 as a biological control agent and consequently, its application in agriculture.

  7. Potential Bio-Control Agent from Rhodomyrtus tomentosa against Listeria monocytogenes

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    Grace Fiyinfoluwa Odedina

    2015-09-01

    Full Text Available Listeria monocytogenes is an important foodborne pathogen implicated in many outbreaks of listeriosis. This study aimed at screening for the potential use of Rhodomyrtus tomentosa ethanolic leaf extract as a bio-control agent against L. monocytogenes. Twenty-two L. monocytogenes isolates were checked with 16 commercial antibiotics and isolates displayed resistance to 10 antibiotics. All the tested isolates were sensitive to the extract with inhibition zones ranging from 14 to 16 mm. Minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC values ranged from 16 to 32 µg/mL and 128 to 512 µg/mL, respectively. Time-kill assay showed that the extract had remarkable bactericidal effects on L. monocytogenes. The extract at a concentration of 16 µg/mL reduced tolerance to 10% NaCl in L. monocytogenes in 4 h. Stationary phase L. monocytogenes cells were rapidly inactivated by greater than 3-log units within 30 min of contact time with R. tomentosa extract at 128 µg/mL. Electron microscopy revealed fragmentary bacteria with changes in the physical and morphological properties. Our study demonstrates the potential of the extract for further development into a bio-control agent in food to prevent the incidence of L. monocytogenes contamination.

  8. Implementing plant biostimulants and biocontrol strategies in the agroecological management of cultivated ecosystems. A review

    Directory of Open Access Journals (Sweden)

    Le Mire, G.

    2016-01-01

    Full Text Available Introduction. In the context of sustainable agricultural production, agroecology aims at optimizing the economic and environmental performances of beneficial ecosystem services in order to (i increase the productivity and resilience of cultivated ecosystems and (ii preserve their natural resources. The maintenance of such performances is supported by research via the development of new tools that enhance plant tolerance to numerous biotic and abiotic stresses. Literature. Biostimulants can be used as a tool to complement the use of chemical inputs, by involving non-living-based products, or living-based products containing beneficial rhizosphere microbiome, such as plant growth-promoting rhizobacteria (PGPR. Pest management research has also made major advances in the development of efficient biocontrol methods. Elicitors and semiochemicals are considered to be some of the most promising tools for inducing plant resistance to various diseases and enhancing natural predation, respectively. Several products are already on the market. This review discusses current methods for exploiting and applying biostimulant and biocontrol products in contemporary agricultural systems. Future applications of these tools for sustainable management of cultivated ecosystems are also discussed. Conclusions. These tools are still difficult to use because of their lack of reliability in the field and their uneasy integration in the cropping systems. Further studies are needed to better understand the parameters influencing the efficiency of PGPR, elicitors and semiochemicals. Special attention needs to be given to the formulation and the interactions of these products with plant physiology and the environment.

  9. Biocontrol mechanism by root-associated Bacillus amyloliquefaciens FZB42 - a review

    Directory of Open Access Journals (Sweden)

    Soumitra ePaul Chowdhury

    2015-07-01

    Full Text Available Bacillus amyloliquefaciens subsp. plantarum FZB42 is a gram-positive model bacterium for unraveling plant-microbe interactions in Bacilli. In addition, FZB42 is used commercially as biofertilizer and biocontrol agent in agriculture. Genome analysis of FZB42 revealed that nearly 10% of the FZB42 genome is devoted to synthesizing antimicrobial metabolites and their corresponding immunity genes. However, recent investigations in planta demonstrated that - except surfactin - the amount of such compounds found in vicinity of plant roots is relatively low, making doubtful a direct function in suppressing competing microflora including plant pathogens. These metabolites have been also suspected to induce changes within the rhizosphere microbial community, which might affect environment and plant health. However, sequence analysis of rhizosphere samples revealed only marginal changes in the root microbiome, suggesting that secondary metabolites are not the key factor in protecting plants from pathogenic microorganisms. On the other hand, adding FZB42 to plants compensate, at least in part, changes in the community structure caused by the pathogen, indicating an interesting mechanism of plant protection by beneficial Bacilli.Sub-lethal concentrations of cyclic lipopeptides and volatiles produced by plant-associated Bacilli trigger pathways of induced systemic resistance (ISR, which protect plants against attacks of pathogenic microbes, viruses and nematodes. Stimulation of ISR by bacterial metabolites is likely the main mechanism responsible for biocontrol action of FZB42.

  10. Relationship between mycoparasites lifestyles and biocontrol behaviors against Fusarium spp. and mycotoxins production.

    Science.gov (United States)

    Kim, Seon Hwa; Vujanovic, Vladimir

    2016-06-01

    Global food security research is seeking eco-friendly solutions to control mycotoxins in grain infected by fungi (molds). In particular, mycotoxigenic Fusarium spp. outbreak is a chronic threat for cereal grain production, human, and animal health. In this review paper, we discuss up-to-date biological control strategies in applying mycoparasites as biological control agents (BCA) to prevent plant diseases in crops and mycotoxins in grain, food, and feed. The aim is to increase food safety and to minimize economic losses due to the reduced grain yield and quality. However, recent papers indicate that the study of the BCA specialists with biotrophic lifestyle lags behind our understanding of the BCA generalists with necrotrophic lifestyle. We examine critical behavioral traits of the two BCA groups of mycoparasites. The goal is to highlight their major characteristics in the context of future research towards an efficient biocontrol strategy against mycotoxin-producing Fusarium species. The emphasis is put on biocontrol of Fusarium graminearum, F. avenaceum, and F. culmorum causing Fusarium head blight (FHB) in cereals and their mycotoxins. PMID:27121573

  11. Bio-control trials of Chaetomium spirale ND35 against apple canker

    Institute of Scientific and Technical Information of China (English)

    XINYa-fen; SHANGJin-jie

    2005-01-01

    A new endophytic antagonistic fungus, Chaetomium spirale ND35 from Populus tomentosa, was reported. The bio-control trials of C. spirale ND35 against the Valsa Canker of apple were preliminarily investigated. The results of dual culture on PDA plate showed that C. spirale ND35 was capable of strong antagonism against Valsa ceratosperma, and for inhibiting the mycelial growth of V. ceratosperma,.the crude extract of liquid culture of corn steep powder broth was more effective than that one of malt extract broth (MEB). The results of bio-control in greenhouse and field indicated that the disease incidence of apple tree treated with C. spirale ND35 was lower significantly than that treated by other methods. The re-isolation experiment suggested that C. spirale ND35 could colonize in stems and branches of apple trees successfully, and the ND35 colonization rate of the treatment with solid wheat bran culture was higher than that of corn steep powder broth, but the field experiment result the control effect of liquid culture of C. spirale ND35 was better than that of solid culture.

  12. Biocontrol activity of Paenibacillus polymyxa AC-1 against Pseudomonas syringae and its interaction with Arabidopsis thaliana.

    Science.gov (United States)

    Hong, Chi Eun; Kwon, Suk Yoon; Park, Jeong Mee

    2016-04-01

    Paenibacillus polymyxa AC-1 (AC-1) is a plant growth-promoting rhizobacterium (PGPR) that has been used as a soil inoculant for biocontrol of plant pathogenic fungi and to promote plant growth. In this study, we examine the effects of AC-1 on the bacterial phytopathogen Pseudomonas syringae and internal colonization of AC-1 by counting bacterial populations that colonize plants. AC-1 inhibited the growth of both P. syringae pv. tomato DC3000 (Pst) and P. syringae pv. tabaci (Pta) in a concentration-dependent manner in in vitro assays. Upon treatment of AC-1 dropping at root tip of axenically grown Arabidopsis, we found that most of the AC-1 was detected in interior of leaves of Arabidiopsis plants rather than roots after 5 days post infection, indicating systemic spreading of AC-1 occur. We examined further AC-1 colonization patterns in Arabidopsis mutants deficient in phytohormone signaling pathways. These results indicated that abscisic acid (ABA) and jasmonic acid (JA) signaling pathways positively and negatively contributed, respectively, to AC-1 colonization of leaves, whereas epiphytic accumulation of AC-1 around root tissues was not affected. This study shows that AC-1 is an effective biocontrol agent to suppress P. syringae growth, possibly owing to its colonization patterns as a leaf-inhabiting endophyte. The results showed in this work will help to expand our understanding of the mode of action of AC-1 as a biological control agent and consequently, its application in agriculture. PMID:26946374

  13. Plant growth-promoting rhizobacteria (PGPR: their potential as antagonists and biocontrol agents

    Directory of Open Access Journals (Sweden)

    Anelise Beneduzi

    2012-01-01

    Full Text Available Bacteria that colonize plant roots and promote plant growth are referred to as plant growth-promoting rhizobacteria (PGPR. PGPR are highly diverse and in this review we focus on rhizobacteria as biocontrol agents. Their effects can occur via local antagonism to soil-borne pathogens or by induction of systemic resistance against pathogens throughout the entire plant. Several substances produced by antagonistic rhizobacteria have been related to pathogen control and indirect promotion of growth in many plants, such as siderophores and antibiotics. Induced systemic resistance (ISR in plants resembles pathogen-induced systemic acquired resistance (SAR under conditions where the inducing bacteria and the challenging pathogen remain spatially separated. Both types of induced resistance render uninfected plant parts more resistant to pathogens in several plant species. Rhizobacteria induce resistance through the salicylic acid-dependent SAR pathway, or require jasmonic acid and ethylene perception from the plant for ISR. Rhizobacteria belonging to the genera Pseudomonas and Bacillus are well known for their antagonistic effects and their ability to trigger ISR. Resistance-inducing and antagonistic rhizobacteria might be useful in formulating new inoculants with combinations of different mechanisms of action, leading to a more efficient use for biocontrol strategies to improve cropping systems.

  14. Regulation of GacA in Pseudomonas chlororaphis Strains Shows a Niche Specificity.

    Directory of Open Access Journals (Sweden)

    Jun Li

    Full Text Available The GacS/GacA two-component system plays a central role in the regulation of a broad range of biological functions in many bacteria. In the biocontrol organism Pseudomonas chlororaphis, the Gac system has been shown to positively control quorum sensing, biofilm formation, and phenazine production, but has an overall negative impact on motility. These studies have been performed with strains originated from the rhizosphere predominantly. To investigate the level of conservation between the GacA regulation of biocontrol-related traits in P. chlororaphis isolates from different habitats, the studies presented here focused on the endophytic isolate G5 of P. chlororaphis subsp. aurantiaca. A gacA mutant deficient in the production of N-acylhomoserine lactones (AHLs and phenazine was isolated through transposon mutagenesis. Further phenotypic characterization revealed that in strain G5, similar to other P. chlororaphis strains, a gacA mutation caused inability to produce biocontrol factors such as phenazine, HCN and proteases responsible for antifungal activity, but overproduced siderophores. LC-MS/MS analysis revealed that AHL production was also practically abolished in this mutant. However, the wild type exhibited an extremely diverse AHL pattern which has never been identified in P. chlororaphis. In contrast to other isolates of this organism, GacA in strain G5 was shown to negatively regulate biofilm formation and oxidative stress response whilst positively regulating cell motility and biosynthesis of indole-3-acetic acid (IAA. To gain a better understanding of the overall impact of GacA in G5, a comparative proteomic analysis was performed revealing that, in addition to some of the traits like phenazine mentioned above, GacA also negatively regulated lipopolysaccharide (LPS and trehalose biosynthesis whilst having a positive impact on energy metabolism, an effect not previously described in P. chlororaphis. Consequently, GacA regulation shows a

  15. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7

    Directory of Open Access Journals (Sweden)

    M. Mercedes eMaldonado-González

    2015-04-01

    Full Text Available The effective management of Verticillium wilts, diseases affecting many crops and caused by some species of the soil-borne fungus Verticillium, is problematic. The use of microbial antagonists to control these pathologies fits modern sustainable agriculture criteria. Pseudomonas fluorescens PICF7 is an endophytic bacterium isolated from olive roots with demonstrated ability to control Verticillium wilt of olive caused by the highly-virulent, defoliating (D pathotype of Verticillium dahliae Kleb. However, the study of the PICF7-V.dahliae-olive tripartite interaction poses difficulties because of the inherent characteristics of woody, long-living plants. To overcome these problems we explored the use of the model plant Arabidopsis thaliana. Results obtained in this study showed that: (i olive D and non-defoliating (ND V. dahliae pathotypes produce differential disease severity in A. thaliana plants; (ii strain PICF7 is able to colonize and persist in the A. thaliana rhizosphere but is not endophytic in Arabidopsis; and (iii strain PICF7 controls Verticillium wilt (VW in Arabidopsis. Additionally, as previously observed in olive, neither swimming motility nor siderophore production by PICF7 are required for VW control in A. thaliana, whilst cysteine auxotrophy decreased the effectiveness of PICF7. Moreover, when applied to the roots PICF7 controlled Botrytis cinerea infection in the leaves of Arabidopsis, suggesting that this strain is able to induce systemic resistance. Arabidopsis thaliana is therefore a suitable alternative to olive bioassays to unravel biocontrol traits involved in biological control of V. dahliae by P. fluorescens PICF7.

  16. Effects of nitrogen availability and spore concentration on the biocontrol activity of Ascochyta caulina in common lambsquarters (Chenopodium album)

    NARCIS (Netherlands)

    Ghorbani, R.; Scheepens, P.C.; Zweerde, van der W.; Leifert, C.; McDonald, A.J.S.; Seel, W.

    2002-01-01

    Common lambsquarters is an annual weed of many important crops. Ascochyta caulina is a plant pathogenic fungus that causes necrotic lesions on the leaves and stems of common lambsquarters. The objective of the present study was to estimate the effect of plant N supply on the biocontrol activity of A

  17. Biocontrol potential of Steinernema thermophilum and its symbiont Xenorhabdus indica against lepidopteran pests: virulence to egg and larval stages

    Science.gov (United States)

    Under laboratory conditions, the biocontrol potential of Steinernema thermophilum was tested against eggs and larval stages of two important lepidopteran insect pests, Helicoverpa armigera and Spodoptera litura (polyphagous pests), as well as Galleria mellonella (used as a model host) . In terms of ...

  18. Biocontrol of Escherichia coli O157:H7 on fresh-cut lettuce and cantaloupe by treatment with bacteriophage

    Science.gov (United States)

    Introduction: Outbreaks of foodborne illness have been associated with the consumption of cantaloupes and fresh-cut lettuce. Bacteriophage mixtures may be effective biocontrol agents to reduce E. coli O157:H7 on produce. Purpose: The effectiveness of a mixture of bacteriophages (ECP-100) in reducin...

  19. Conidial quality of the biocontrol agent Coniothyrium minitans produced by solid-state cultivation in a packed-bed reactor

    NARCIS (Netherlands)

    Jones, E.E.; Weber, F.J.; Oostra, J.; Rinzema, A.; Mead, A.; Whipps, J.M.

    2004-01-01

    Conidial germination and novel sand- and soil-based sclerotial parasitism assays were used to test the quality of conidial inoculum of the biocontrol agent Coniothyrium minitans IVT1 produced in different packed bed reactor runs. The fermenter airflow rate was either kept constant, resulting in a tr

  20. Exploiting the genetic diversity of Beauveria bassiana for improving the biological control of the coffee berry borer through the use of strain mixtures.

    Science.gov (United States)

    Cruz, Lina P; Gaitan, Alvaro L; Gongora, Carmenza E

    2006-08-01

    Beauveria bassiana is an entomopathogen widely used to control the coffee berry borer in Colombia, as part of an Integrated Pest Management strategy. Traditionally, the development of fungal insect pathogens as biocontrol agents in crop pests has been oriented towards the selection and formulation of elite clonal strains. Instead, we explored the potential application of genetic diversity in B. bassiana by determining the effect of strain mixtures on coffee berry borer mortality compared to clonal isolates. Genomic DNA from 11 strains was characterized using internal transcribed spacers and beta-tubulin sequences as well as amplified fragment length polymorphism markers. Cluster analysis produced three genetic groups and confirmed the low but significant intraspecific genetic diversity present among the strains. Single strain virulence towards the coffee berry borer under laboratory conditions, using 1x10(6) conidia ml(-1), ranged between 89.9 and 57.5%. All the inoculations with mixtures resulted in coinfection events. Combinations of genetically similar strains showed no significant differences when their virulences were compared. However, mixtures of genetically different strains led to both antagonism and synergism. The lowest virulence percentage (57%) was obtained by putting together the most virulent strain of each group, contrary to the highest virulence percentage (93%) that resulted from mixing the three least virulent strains. The results indicate the promising potential of designing strain mixtures as an alternative for the biocontrol of Hypothenemus hampei and other pests and provide tools for the understanding of the ecological dynamics of entomopathogen populations under natural conditions. PMID:16362818

  1. 生防解淀粉芽胞杆菌(Bacillus amyloliquefaciens)最新研究进展%The Latest Advances in Bio-Control Bacillus amyloliquefaciens

    Institute of Scientific and Technical Information of China (English)

    李红晓; 张殿朋; 卢彩鸽; 赵洪新

    2016-01-01

    Bacillus amyloliquefaciens is a strain similar to B. subtilis,a beneficial microbe that can produce spore, Gram-positive,and possessing biocontrolling activity. During its metabolism it produces antibiotic substance,a varie-ty of antibiotic preparations or antibiotic extracts with broad antibiotic spectrum,non toxic,non harms,non residues, long effectiveness for a given period of time,and other advantages. It can be widely applied in keeping fruits and vegetables fresh,and in biocontrolling microbial pests during their growth and post-harvest. It is a very ideal biocon-trolling microbe with the most developing value. In this paper the studies,the current application of B. amyloliquefa-ciens were drawn and summarized from aspect of the antibiotic mechanism,the antibiotic gene component,the estab-lishment of genetic engineering strain,the development of resistance products,the controlling against microbial pests,and other aspects,and accordingly to provide references for its large-scale safety application in agricultural bio-engineering.%解淀粉芽胞杆菌(Bacillus amyloliquefaciens)是与枯草芽胞杆菌相似,可产芽胞、革兰阳性(G +)、具有生防活性的益生细菌。其代谢过程中产生的抗菌物质种类多、抗菌谱广,其抗菌制剂或抗菌提取物具有无毒、无害、无残留、抑菌时效长等优点,可广泛应用于果蔬保鲜、果蔬生长期及采后微生物病害的生物防治,是极具开发价值的生防菌。本文从抗菌机理、抗性基因组成、基因工程菌构建、抗性产品开发及对微生物病害防治等几个方面,对解淀粉芽胞杆菌的研究及应用现状进行了归纳和总结,为其在农业生物工程中的规模化安全应用提供参考。

  2. Production and characterization of rhamnolipids produced by Serratia rubidaea SNAU02 under solid-state fermentation and its application as biocontrol agent.

    Science.gov (United States)

    Nalini, S; Parthasarathi, R

    2014-12-01

    The present study aimed at exploring mahua (Madhuca indica) oil cake as a novel substrate for the production of biosurfactant by Serratia rubidaea SNAU02 under solid-state fermentation (SSF). Response surface methodology showed followings as the optimal conditions for the production of biosurfactant: mahua oil cake 7.48 g, 2.5 ml inoculum size (1×10(8) cells/ml), and pH 7.22 and 31 °C temperature. The characterization of the biosurfactant by TLC, FT-IR and GC-MS revealed the presence of rhamnolipid. The presence of rhamnosyl transferase gene responsible for biosynthesis of rhamnolipid was identified. The strain SNAU02 exhibited antifungal activity and demonstrated no toxicity against the seeds of Brassica oleracea and Artemia salina employed as a bio-indicator. The present findings indicated the potential of mahua oil cake as suitable substrate for the production of rhamnolipids in SSF by S. rubidaea SNAU02 and application potential of the biosurfactant produced as biocontrol agent against plant pathogens. PMID:25305653

  3. Isolation of Stem rot Disease Causing Organism of Brinjal and their in-vitro Inhibition with Fungicides and Bio-control Agents

    Directory of Open Access Journals (Sweden)

    Shaily Javeria

    2014-09-01

    Full Text Available Different strains of Sclerotinia sclerotiorum were isolated from the diseased samples collected from different hosts and locations. Among the 14 isolates, 12 isolates colonies covered the entire Petri plates within 96 hours but, two isolates from fababean and yellow mustard showed slow colony growth within 96 hours. All isolates produced sclerotia which were varied in number, but the fenugreek isolate produced maximum (43 number of sclerotia and lambs quarter isolate produced minimum number of sclerotia (12 on PDA medium. To examine inhibitory effect of fungicide on the mycelial growth of the pathogen, 9 fungicides were tested in vitro against Sclerotinia sclerotiorum, of those carbendazim, carboxin, topsin-M and carbendazim+ mancozeb (SAAF were found most effective and inhibited the mycelial growth of pathogen up to 100 per cent at 0.05%, 0.1%, and 0.2% concentration. The effect of different bioagents viz., Trichoderma harzianum, T. viride, T. koningii, T. atroviride, T. longibraciatum, Aspergillus niger, Chaetomium globosome and Penicillium notatum in inhibiting the growth of Sclerotinia sclerotiorum was studied through “Dual Culture Technique”. The data showed that among the eight biocontrol agent six were fond effective. The maximum inhibition was found by T. harzianum causing 70.82% inhibition of mycelial growth of the pathogen S. sclerotiorum.

  4. Integrated effect of gamma radiation and biocontrol agent on quality parameters of apple fruit: An innovative commercial preservation method

    International Nuclear Information System (INIS)

    Effects of gamma irradiation and biocontrol agent (Pseudomonas fluorescens) on the physico-chemical parameters (including moisture, total soluble solids, antioxidant activity, phenolic content and firmness) of cv. Golden Delicious apples were investigated for their ability to avoid the post-harvest blue mold caused by Penicillium expansum during cold storage. Freshly harvested apples were inoculated with P. expansum. Treated fruits were irradiated at doses of 0, 200, 400, 600 and 800 Gy and then inoculated with P. fluorescens suspension. Samples were evaluated at 3 month intervals. The results demonstrated a clear link between antioxidant activity and phenolic content, so that dose range of 200–400 Gy significantly increased phenolic content and antioxidant activity. Effect of P. fluorescens was similar to irradiation at 200 and 400 Gy that could prevent lesion diameter in pathogen-treated apples. As dose and storage time increased firmness decreased but, combination of P. fluorescens as well as irradiation (at 200–400 Gy) could decrease softening apple fruits during storage. In all parameters, P. fluorescens (as biocontrol agent) inhibited P. expansum similar to irradiation at 200–400 Gy. So, integrated treatment of irradiation and biocontrol agent explored the potential dual benefit of low doses (200 and 400 Gy) as a suitable method to sustain physico-chemical quality and conclusively reduce apple fruits losses during post-harvest preservation. - Highlights: • A suitable method to reduce apple quality losses during 9 month storage period. • Effects of γ radiation in combination with biocontrol agent on physico-chemical parameters of the apple fruits during cold storage. • The potential dual benefit of low irradiation dose combined with biocontrol agent. • Radiation dose determination for Penicillium expansum (postharvest pathogen) control

  5. 生防细菌SF-16产芽胞条件的研究%Studies on Biocontrol Bacteria SF-16 Spore Production Condition

    Institute of Scientific and Technical Information of China (English)

    张明; 李欢; 朱莹

    2012-01-01

    生防细菌SF-16为土壤中筛选得到的1株细菌,研究表明,其对多种植物致病真菌有抑制作用。为了提高生防细菌SF-16的芽胞形成率及芽胞数量,以及为工厂化生产生物菌剂提供理论依据,采用摇瓶发酵对芽胞形成的主要影响因素进行了考察。通过单因素实验和正交实验对SF-16菌株产芽胞条件进行了分析,确定了摇瓶发酵最佳条件为:乳糖0.5%、黄豆饼粉2.0%、MnSO4·5H2O0.30%。在该优化条件下,发酵液中颉颃菌的出芽率达到98.3%,最终生物量达到2×109个/mL。%Biocontrol bacteria SF-16 was a selected bacteria strain,research showed that its inhibitory effect on multiple plants.In order to improve biocontrol bacteria SF-16 spore formation rate and quantity,provide theoretical basis for factory production of biological agent,use flask fermentation with a survey on main influencing factors of spore formation.Through single factor experiment and orthogonal experiment,the optimal shaking flask fermentation condition was determined as follow:media composed of 0.5% lactose,2.0% soybean meal,0.30% MnSO4·5H2O.Under this optimum condition,the germination rate achieves 98.3 percentages,Biomass achieves 2×109/mL.

  6. Wonder world of phages: potential biocontrol agents safeguarding biosphere and health of animals and humans- current scenario and perspectives.

    Science.gov (United States)

    Tiwari, Ruchi; Chakraborty, Sandip; Dhama, Kuldeep; Wani, Mohd Yaqoob; Kumar, Amit; Kapoor, Sanjay

    2014-02-01

    Darwin's theory of natural selection and concept of survival of fittest of Wallace is a universal truth which derives the force of life among all live entities on this biosphere. Issues regarding food safety along with increased drug resistance and emerging zoonotic infections have proved that multidisciplinary efforts are in demand for human and animal welfare. This has led to development of various novel therapies the list of which remains incomplete without mentioning about phages. Homologous and non-homologous recombination along with point mutation and addition of new genes play role in their evolution. The rapid emergence of the antibiotic resistant strains of bacteria have created keen interest in finding necessary alternatives to check microbial infections and there comes the importance of phages. Phages kill the bacteria either by lysis or by releasing holins. Bacteriophages; the viruses that live on bacteria are nowadays considered as the best biocontrol agents. They are used as replacers of antibiotics; food industry promoter; guard of aquatic life as well as of plants; pre-slaughter treatment agents; Generally Recognized As Safe (GRAS) food additives; Typing agent of bacteria; active tool of super bug therapy; in post harvest crops and food and during post infection and also to combat intracellular pathogens viz. Mycobacteria and Mycoplasma. Cyanophages/phycophages are particularly useful in controlling blooms produced by various genera of algae and cyanobacteria. By performing centrifugation studies and based on electron microscopy certain virus like particles containing ds RNA have been confirmed as mycophages. They are well proven as threat to pathogenic fungi (both fungal hyphae and yeast). Those that infect yeasts are called zymophages. Virophages have exquisite specificity for their viral host, hence can extensively be used for genetic studies and can also act as evolutionary link. After the discovery of very first virophage till now, a total of 3

  7. Production of trichodiene by Trichoderma harzianum alters the perception of this biocontrol strain by plants and antagonized fungi

    Science.gov (United States)

    Trichothecenes are phytotoxic sesquiterpenoid compounds of fungal origin which can act as virulence factors in plant diseases. Harzianum A (HA) is a non-phytotoxic trichothecene produced by Trichoderma arundinaceum. The first step in the biosynthesis of HA is the conversion of farnesyl diphosphate t...

  8. Genome Sequence of the Endophytic Bacterium Bacillus thuringiensis Strain KB1, a Potential Biocontrol Agent against Phytopathogens.

    Science.gov (United States)

    Jeong, Haeyoung; Jo, Sung Hee; Hong, Chi Eun; Park, Jeong Mee

    2016-04-21

    ITALIC! Bacillus thuringiensisis the most widely known microbial pesticide used in agricultural applications. Herein, we report a draft genome sequence of the endophytic bacterium ITALIC! Bacillus thuringiensisstrain KB1, which exhibits antagonism against phytopathogens.

  9. Construction of an environmental safe Bacillus thuringiensis engineered strain against Coleoptera.

    Science.gov (United States)

    Yu, Yajun; Yuan, Yihui; Gao, Meiying

    2016-05-01

    Cloning of new toxic genes from Bacillus thuringiensis (Bt) and construction of Bt engineered strains are two key strategies for bio-control of coleopteran pests in agriculture and forestry. In this study, we cloned a new cry3Aa-type gene, cry3Aa8, from wild Bt strain YC-03 against coleopteran, and constructed a Bt engineered strain, ACE-38, containing insecticidal protein-encoding gene cry3Aa8. The engineered strain, with almost four times of Cry3Aa yield compared with strain YC-03, was an antibiotic marker-free strain. Though no selective pressure was presented in the medium, cry3Aa8 in the engineered strain ACE-38 remained stable. The yield of Cry3Aa by strain ACE-38 reached 2.09 mg/ml in the optimized fermentation medium. The activity of strain ACE-38 against Plagiodera versicolora was tested, and the LC50 of ACE-38 cultures in the optimized fermentation medium was 1.13 μl/ml. Strain ACE-38 is a non-antibiotic Bt engineered strain with high Chrysomelidae toxicity and exhibits good fermentation property. The modified indigenous site-specific recombination system constructed in this study might be useful for the construction of Bt engineered strains containing genes that cannot be expressed in the indigenous site-specific recombination system using plasmid pBMB1205R. PMID:26767987

  10. Construction of an environmental safe Bacillus thuringiensis engineered strain against Coleoptera.

    Science.gov (United States)

    Yu, Yajun; Yuan, Yihui; Gao, Meiying

    2016-05-01

    Cloning of new toxic genes from Bacillus thuringiensis (Bt) and construction of Bt engineered strains are two key strategies for bio-control of coleopteran pests in agriculture and forestry. In this study, we cloned a new cry3Aa-type gene, cry3Aa8, from wild Bt strain YC-03 against coleopteran, and constructed a Bt engineered strain, ACE-38, containing insecticidal protein-encoding gene cry3Aa8. The engineered strain, with almost four times of Cry3Aa yield compared with strain YC-03, was an antibiotic marker-free strain. Though no selective pressure was presented in the medium, cry3Aa8 in the engineered strain ACE-38 remained stable. The yield of Cry3Aa by strain ACE-38 reached 2.09 mg/ml in the optimized fermentation medium. The activity of strain ACE-38 against Plagiodera versicolora was tested, and the LC50 of ACE-38 cultures in the optimized fermentation medium was 1.13 μl/ml. Strain ACE-38 is a non-antibiotic Bt engineered strain with high Chrysomelidae toxicity and exhibits good fermentation property. The modified indigenous site-specific recombination system constructed in this study might be useful for the construction of Bt engineered strains containing genes that cannot be expressed in the indigenous site-specific recombination system using plasmid pBMB1205R.

  11. Potencialidade de um actinomiceto de rizosfera de tomateiro como agente de biocontrole de doenças Potenciality of an actinomycete from tomato rhizosphere as a biocontrol agent for tomato diseases

    Directory of Open Access Journals (Sweden)

    Renato Carrer Filho

    2009-09-01

    Full Text Available Um actinomiceto (Streptomyces setonii, isolado 'UFV-RD1', obtido de rizosfera de planta sadia de tomateiro, foi selecionado dentre outros 117, como promissor agente de biocontrole de enfermidades da cultura. Em testes de antagonismo in vitro contra patógenos do tomateiro, o isolado 'UFV-RD1' foi incapaz de inibir o crescimento de bactérias (Pseudomonas syringae pv. tomato, Ralstonia solanacearum, Pectobacterium carotovorum subsp. carotovorum, Xanthomonas campestris pv. vesicatoria mas inibiu a germinação de conídios de alguns fungos (Alternaria solani, Phytophthora infestans, Corynespora cassiicola, Stemphylium solani. Em ensaios de biocontrole experimental in vivo, em casa de vegetação, o actinomiceto foi efetivo em reduzir a severidade de sintomas no caso de patógenos fúngicos e bacterianos testados como desafiantes. A campo, quando A. solani e P. infestans ocorreram naturalmente, as plantas originárias de sementes microbiolizadas com propágulos da estirpe 'UFV-RD1' exibiram sintomas menos severos que as plantas controle para o caso da pinta preta. O agente de biocontrole é promissor para futuros protocolos de manejo integrado, como forma de reduzir a quantidade de defensivos utilizados.An actinomycete (Streptomyces setonii, isolate 'UFV-RD1', isolated from the rhizosphere of a healthy tomato plant was selected out of 117 as a promising biocontrol agent for tomato diseases. In in vitro antagonism tests against tomato pathogens, the isolate 'UFV-RD1' was unable to inhibit growth of bacterial pathogens (Pseudomonas syringae pv. tomato, Ralstonia solanacearum, Pectobacterium carotovorum subsp. carotovorum, Xanthomonas campestris pv. vesicatoria but inhibited conidium germination of fungi (Alternaria solani, Phytophthora infestans, Corynespora cassiicola, Stemphylium solani. Experimental biocontrol assays in a greenhouse indicated that the actinomycete was effective for reducing symptom severity in the case of bacteria and fungi tested

  12. The biocontrol efficiency and identification of cyclamen soft rot antagonistic bacteria%仙客来软腐病拮抗细菌鉴定及其生物防治效果

    Institute of Scientific and Technical Information of China (English)

    钱仁卷; 姚丽娟; 刘洪见; 张旭乐; 黄建; 凌树椹; 林志秀; 刘佳明

    2012-01-01

    Objective To select Cyclamen soft rot bacteria and evaluatt its biocontrol effect. Method Thirteen antagonistic strains against Erwinia spp were separated from healthy cyclamen plants collected from Wenzhou Zeya Mountain. Among them the strain Yl possessed prominent bioactivity and genetic stabilization. Its morphology, physiological and biochemical properties, 16S rDNA and phylogenetic tree were studied and analyzed. Result The strain Yl was identified as Bacillus spp. Its fermenting liquid was used onto normal plants, and the results showed that the soft rot incidence of Cyclamen treated by the cell culture of strain Yl were more than 60%. Conclusion The strain Yl have the characteristics of biocontrol, which could have an application prospect in the flower industry.%目的 为筛选出仙客来软腐病拮抗细菌及评价其生防效果.方法 通过分离和筛选,从温州泽雅高山基地采集健康的仙客来植株分离到13株内生细菌对仙客来软腐病病原菌有较强的抑制作用,其中菌株Y1活性最强且遗传稳定.通过形态特征、生理生化特性分析、16S rDNA序列测定及其系统发育分析研究.结果 菌株Y1鉴定为芽胞杆菌,将菌株Y1以发酵液灌根方式回接正常仙客来植株,菌株Y1的发酵液处理仙客来软腐病的防效均大于60%以上.结论 菌株Y1具有生防的特点,在花卉产业上具有应用潜力.

  13. Shallot aphids, Myzus ascalonicus, in strawberry: biocontrol potential of three predators and three parasitoids.

    Science.gov (United States)

    Enkegaard, Annie; Sigsgaard, Lene; Kristensen, Kristian

    2013-01-01

    The parasitization capacity of 3 parasitoids and the predation capacity of 3 predators towards the shallot aphid, Myzus ascalonicus Doncaster (Homoptera: Aphididae), on strawberry, Fragaria x ananassa Duchesne (Rosales: Rosaceae) cv. Honeoye, were examined in laboratory experiments. In Petri dish assays, both Aphidius colemani Viereck (Hymenoptera: Aphidiidae) and A. ervi Haliday readily stung shallot aphids, with no significant difference in stinging frequency between the two species. A. ervi induced a significantly higher mortality (79.0 ± 7.2%) in terms of stung aphids compared with A. colemani (55.3 ± 4.1%); however, only a minor fraction (2.7 ± 1.8% and 7.1 ± 3.1%, respectively) of the killed aphids resulted in formation of mummies, presumably due to a physiological response to parasitism. The low percentage of mummification precludes the use of either Aphidius species in anything but inundative biocontrol. In similar set-ups, Aphelinus abdominalis (Dalman) (Hymenoptera: Aphelinidae) killed almost half (49.6 ± 5.3%) of the exposed aphids through host feeding. In addition, 23.2 ± 7.3% of non-host-fed aphids developed into mummified aphids, and 38.1 ± 13.2% of non-host-fed aphids died from other parasitoid-induced causes. However, the host feeding rate was reduced to only 1.2 ± 0.8%, and no significant parasitization mortality was observed on strawberry plants, suggesting that host plants interfered with A. abdominalis activity. This parasitoid does not, therefore, seem to be suited to either inoculative or inundative biocontrol of shallot aphids in strawberry. The three predators studied were the green lacewing, Chrysoperla carnea Steph. (Neuroptera: Chrysopidae), the two-spotted lady beetle, Adalia bipunctata L. (Coleoptera: Coccinellidae), and the gall midge Aphidoletes aphidimyza (Rondani) (Diptera: Cecidomyiidae). Third instars of all 3 predators readily preyed upon the shallot aphid in Petri dish set-ups with significant differences in daily

  14. EFFICACY OF BIOCONTROL AGENTS IN CONTROLLING RHIZOCTONIA SOLANI ON NAGA KING CHILLI ( Capsicum chinense Jacq.

    Directory of Open Access Journals (Sweden)

    Marinus Ngullie

    2013-07-01

    Full Text Available Available biocontrol agents were evaluated either alone or in various combinations for finding out their efficacy in suppressing Rhizoctonia seedling rot incidence and promoting plant growth of Naga king chilli (Capsicum chinense Jacq. in green house as well field conditions. Among all tested combination, the treatment containing combination of T. viride +P. fluorescens was found most effective in reducing the incidence of seedling rot in both greenhouse and field condition. Highest per plant yield was also recorded from the same combination and it was followed by T. viride. However, the commercially available fungicide Copper oxychloride (0.1% showed 9.82 % and11.88% disease incidence in greenhouse and field condition respectively

  15. Synthesis of biocontrol macromolecules by derivative of chitosan with surfactin and antifungal evaluation.

    Science.gov (United States)

    Yuan, Bo; Xu, Pei-Yuan; Zhang, Yue-Ji; Wang, Pei-Pei; Yu, Hong; Jiang, Ji-Hong

    2014-05-01

    A derivative of chitosan was prepared with chitosan and β-cyclodextrins, which was synthesized by the immobilization reaction, as a carrier to adsorb surfactin produced from Bacillus amyloliquefaciens and got biological macromolecules. The antifungal activity against three sapstain fungi by a combination of macromolecules was tested. The results showed that the macromolecules inhibited the mycelium growth of sapstain fungi Lasiodiplodia rubropurpurea, L. crassispora, and L. theobromae by about 73.22%, 76.72%, and 70.22%, respectively. The macromolecules were relatively thermally stable with more than 50% of the antifungal activity even after being held at 121°C for 30 min. Meanwhile, the activity of the macromolecules remained more than 55% at a pH value ranging from 4 to 12. The macromolecules were resistant to hydrolysis by most protein-denaturing detergents and other enzymes. The results indicated the macromolecules might provide an alternative bioresource for the bio-control of sapstain. PMID:24530369

  16. Native isolate of Trichoderma: a biocontrol agent with unique stress tolerance properties.

    Science.gov (United States)

    Mishra, N; Khan, S S; Sundari, S Krishna

    2016-08-01

    Species of Trichoderma are widely recognized for their biocontrol abilities, but seldom studied collectively, for their plant growth promotion, abiotic stress tolerance and bioremediation properties. Our study is a concentrated effort to establish the potential of native isolate Trichoderma harzianum KSNM (T103) to tolerate biotic (root pathogens) and abiotic stresses [high salt (100-1000 mM); heavy metal (chromium, nickel and zinc: 1-10 mM); pesticides: malathion (100-600 ppm), carbofuran (100-600 ppb)], along with its ability to support plant growth. In vitro growth promotion assays with T103 treated Vigna radiata, Vigna mungo and Hordeum vulgare confirmed 'non-species specific' growth promotion effects of T103. At lower metal concentration, T103 treatment was found to completely negate the impact of metal stress [60 % increase in radicle length (RL) with no significant decrease in %germination (%G)]. Even at 10 mM metal, T103 inoculation gave 80 % increase in %G and >50 % increase in RL. In vitro experiments confirmed high metal reduction capacity (47 %-Cr, 35 %-Ni and 42 %-Zn) of T103 at concentrations as high as 4 mM. At maximum residual concentrations of malathion (440 ppm) and carbofuran (100 ppb) reported in agricultural soils, T103 maintained 80 and 100 % survivability, respectively. T103 treatment has improved %G and RL in all three hosts challenged with pesticide. Isolate T103 was found to effectively suppress growth of three major root pathogens: Macrophomina phaseolina (65.83 %) followed by Sclerotium rolfsii (19.33 %) and Fusarium oxysporum (19.18 %). In the light of these observations, native T. harzianum (T103) seems to be a competent biocontrol agent for tropical agricultural soils contaminated with residual pesticides and heavy metals. PMID:27339311

  17. The importance of chorismate mutase in the biocontrol potential of Trichoderma parareesei.

    Science.gov (United States)

    Pérez, Esclaudys; Rubio, M Belén; Cardoza, Rosa E; Gutiérrez, Santiago; Bettiol, Wagner; Monte, Enrique; Hermosa, Rosa

    2015-01-01

    Species of Trichoderma exert direct biocontrol activity against soil-borne plant pathogens due to their ability to compete for nutrients and to inhibit or kill their targets through the production of antibiotics and/or hydrolytic enzymes. In addition to these abilities, Trichoderma spp. have beneficial effects for plants, including the stimulation of defenses and the promotion of growth. Here we study the role in biocontrol of the T. parareesei Tparo7 gene, encoding a chorismate mutase (CM), a shikimate pathway branch point leading to the production of aromatic amino acids, which are not only essential components of protein synthesis but also the precursors of a wide range of secondary metabolites. We isolated T. parareesei transformants with the Tparo7 gene silenced. Compared with the wild-type, decreased levels of Tparo7 expression in the silenced transformants were accompanied by reduced CM activity, lower growth rates on different culture media, and reduced mycoparasitic behavior against the phytopathogenic fungi Rhizoctonia solani, Fusarium oxysporum and Botrytis cinerea in dual cultures. By contrast, higher amounts of the aromatic metabolites tyrosol, 2-phenylethanol and salicylic acid were detected in supernatants from the silenced transformants, which were able to inhibit the growth of F. oxysporum and B. cinerea. In in vitro plant assays, Tparo7-silenced transformants also showed a reduced capacity to colonize tomato roots. The effect of Tparo7-silencing on tomato plant responses was examined in greenhouse assays. The growth of plants colonized by the silenced transformants was reduced and the plants exhibited an increased susceptibility to B. cinerea in comparison with the responses observed for control plants. In addition, the plants turned yellowish and were defective in jasmonic acid- and ethylene-regulated signaling pathways which was seen by expression analysis of lipoxygenase 1 (LOX1), ethylene-insensitive protein 2 (EIN2) and pathogenesis

  18. The importance of chorismate mutase in the biocontrol potential of Trichoderma parareesei

    Directory of Open Access Journals (Sweden)

    Esclaudys ePérez

    2015-10-01

    Full Text Available Species of Trichoderma exert direct biocontrol activity against soil-borne plant pathogens due to their ability to compete for nutrients and to inhibit or kill their targets through the production of antibiotics and/or hydrolytic enzymes. In addition to these abilities, Trichoderma spp. have beneficial effects for plants, including the stimulation of defenses and the promotion of growth. Here we study the role in biocontrol of the T. parareesei Tparo7 gene, encoding a chorismate mutase (CM, a shikimate pathway branch point leading to the production of aromatic amino acids, which are not only essential components of protein synthesis but also the precursors of a wide range of secondary metabolites. We isolated T. parareesei transformants with the Tparo7 gene silenced. Compared with the wild-type, decreased levels of Tparo7 expression in the silenced transformants were accompanied by reduced CM activity, lower growth rates on different culture media, and reduced mycoparasitic behavior against the phytopathogenic fungi Rhizoctonia solani, Fusarium oxysporum and Botrytis cinerea in dual cultures. By contrast, higher amounts of the aromatic metabolites tyrosol, 2-phenylethanol and salicylic acid were detected in supernatants from the silenced transformants, which were able to inhibit the growth of F. oxysporum and B. cinerea. In in vitro plant assays, Tparo7-silenced transformants also showed a reduced capacity to colonize tomato roots. The effect of Tparo7-silencing on tomato plant responses was examined in greenhouse assays. The growth of plants colonized by the silenced transformants was reduced and the plants exhibited an increased susceptibility to B. cinerea in comparison with the responses observed for control plants. In addition, the plants turned yellowish and were defective in jasmonic acid- and ethylene-regulated signaling pathways which was seen by expression analysis of lipoxygenase 1 (LOX1, ethylene-insensitive protein 2 (EIN2 and

  19. IDIOSYNCRASY OF LOCAL FUNGAL ISOLATE HYPOCREA RUFA STRAIN P2: PLANT GROWTH PROMOTION AND MYCOPARASITISM

    Directory of Open Access Journals (Sweden)

    Parth Thakor

    2016-06-01

    Full Text Available Trichoderma viride an anamorph of Hypocrea rufa, is a known bio-control agent against various fungal phytopathogens. In the present study, H. rufa strain P2 was tested for plant growth promoting (PGP traits and antifungal activity against Fusarium oxysporum, Alternaria alternata, Aspergillus niger, Sclerotium rolfsii. In-vitro assessment of H. rufa strain P2 showed maximum IAA production of 68 µg ml-1, solubilised tri-calcium phosphate up to 72 µg ml-1 and showed production of chitinase enzyme 120 U ml-1. In order to determine in-vivo plant growth promotion, talc based formulation of H. rufa strain P2 was prepared and tested on Arachis hypogaea L. using seed and soil application. After 15 days, treated plants showed six-fold increases in the fresh and dry root mass whereas, fresh and dry shoot mass was increased up to two folds. The result indicates the local isolate H. rufa strain P2 can be categorized as phyto-friendly fungi which can be used as both, bio-control agent as well as phyto-augmenting bio-fertilizer.

  20. Draft Genome Sequence of Plant-Growth-Promoting Rhizobacterium Serratia fonticola Strain AU-AP2C, Isolated from the Pea Rhizosphere

    OpenAIRE

    Devi, Usha; Khatri, Indu; Kumar, Navinder; Sharma, Deepak; Subramanian, Srikrishna; Saini, Adesh K.

    2013-01-01

    Plant health can be augmented by plant-growth-promoting rhizobacteria (PGPR) that confer biofertilizer, phytostimulation, and biocontrol activities. Herein, we provide the high-quality draft genome sequence of Serratia fonticola strain AU-AP2C, a Gram-negative motile PGPR of the pea plant, conferring phosphate solubilization, ammonia production, and antifungal activity against Fusarium sp. The 4.9-Mb genome contains genes related to plant growth promotion and synthesis of siderophores.

  1. Identification and evaluation of strain B37 of Bacillus subtilis antagonistic to sapstain fungi on poplar wood.

    Science.gov (United States)

    Zhang, XiaoHua; Zhao, GuiHua; Li, DeWei; Li, ShunPeng; Hong, Qing

    2014-01-01

    Devaluation of poplar products by sapstain accounts for huge and unpredictable losses each year in China. We had isolated four poplar sapstain fungi, Ceratocystis adiposa Hz91, Lasiodiplodia theobromae YM0737, L. theobromae Fx46, and Fusarium sp. YM05, from five poplar varieties and 13 antagonistic bacteria from nine diverse varieties. After being experimented with agar plates, wood chips, and enzyme activities, strain B37 was identified as the best poplar sapstain biocontrol bacterium. The strain B37 was identified as Bacillus subtilis using sequences of the 16S rRNA gene, physiological biochemical, and morphological characteristics. PMID:25401124

  2. Identification and Evaluation of Strain B37 of Bacillus subtilis Antagonistic to Sapstain Fungi on Poplar Wood

    Directory of Open Access Journals (Sweden)

    XiaoHua Zhang

    2014-01-01

    Full Text Available Devaluation of poplar products by sapstain accounts for huge and unpredictable losses each year in China. We had isolated four poplar sapstain fungi, Ceratocystis adiposa Hz91, Lasiodiplodia theobromae YM0737, L. theobromae Fx46, and Fusarium sp. YM05, from five poplar varieties and 13 antagonistic bacteria from nine diverse varieties. After being experimented with agar plates, wood chips, and enzyme activities, strain B37 was identified as the best poplar sapstain biocontrol bacterium. The strain B37 was identified as Bacillus subtilis using sequences of the 16S rRNA gene, physiological biochemical, and morphological characteristics.

  3. Biocontrol Ability and Action Mechanism of Starmerella bacillaris (Synonym Candida zemplinina) Isolated from Wine Musts against Gray Mold Disease Agent Botrytis cinerea on Grape and Their Effects on Alcoholic Fermentation.

    Science.gov (United States)

    Lemos, Wilson J; Bovo, Barbara; Nadai, Chiara; Crosato, Giulia; Carlot, Milena; Favaron, Francesco; Giacomini, Alessio; Corich, Viviana

    2016-01-01

    Gray mold is one of the most important diseases of grapevine in temperate climates. This plant pathogen affects plant growth and reduces wine quality. The use of yeasts as biocontrol agents to apply in the vineyard have been investigated in recent years as an alternative to agrochemicals. In this work, fermenting musts obtained from overripe grape berries, therefore more susceptible to infection by fungal pathogens such as Botrytis cinerea, were considered for the selection of yeasts carrying antifungal activity. Thirty-six isolates were identified as Starmerella bacillaris, a species recently proven to be of enological interest. Among them 14 different strains were studied and antifungal activity against B. cinerea was demonstrated, for the first time, to be present in S. bacillaris species. The production of volatile organic compounds (VOCs), tested in vitro, was found to be the main responsible of S. bacillaris antifungal effects. All the strains were able to reduce B. cinerea decay on wounded grape berries artificially inoculated with gray mold. The colonization level of wound was very high reaching, after 5 days, a concentration of 10(6) cells per ml of grape juice obtained after berry crushing. At this cell concentration S. bacillaris strains were used to ferment synthetic and natural musts. The sequential yeast inoculation, performed by adding S. cerevisiae 48 h after S. bacillaris, was needed to complete sugar consumption and determined a significant increase in glicerol content and a reduction of ethanol and acetic acid concentrations. The high wound colonization ability, found in this work, together with the propensity to colonize grape berry and the interesting enological traits possessed by the selected S. bacillaris strains allow the use of this yeast as biocontrol agent on vine and grape berries with possible positive effects on must fermentation, although the presence of S. cerevisiae is needed to complete the fermentation process. This work

  4. Biocontrol Ability and Action Mechanism of Starmerella bacillaris (Synonym Candida zemplinina) Isolated from Wine Musts against Gray Mold Disease Agent Botrytis cinerea on Grape and Their Effects on Alcoholic Fermentation

    Science.gov (United States)

    Lemos Junior, Wilson José Fernandes; Bovo, Barbara; Nadai, Chiara; Crosato, Giulia; Carlot, Milena; Favaron, Francesco; Giacomini, Alessio; Corich, Viviana

    2016-01-01

    Gray mold is one of the most important diseases of grapevine in temperate climates. This plant pathogen affects plant growth and reduces wine quality. The use of yeasts as biocontrol agents to apply in the vineyard have been investigated in recent years as an alternative to agrochemicals. In this work, fermenting musts obtained from overripe grape berries, therefore more susceptible to infection by fungal pathogens such as Botrytis cinerea, were considered for the selection of yeasts carrying antifungal activity. Thirty-six isolates were identified as Starmerella bacillaris, a species recently proven to be of enological interest. Among them 14 different strains were studied and antifungal activity against B. cinerea was demonstrated, for the first time, to be present in S. bacillaris species. The production of volatile organic compounds (VOCs), tested in vitro, was found to be the main responsible of S. bacillaris antifungal effects. All the strains were able to reduce B. cinerea decay on wounded grape berries artificially inoculated with gray mold. The colonization level of wound was very high reaching, after 5 days, a concentration of 106 cells per ml of grape juice obtained after berry crushing. At this cell concentration S. bacillaris strains were used to ferment synthetic and natural musts. The sequential yeast inoculation, performed by adding S. cerevisiae 48 h after S. bacillaris, was needed to complete sugar consumption and determined a significant increase in glicerol content and a reduction of ethanol and acetic acid concentrations. The high wound colonization ability, found in this work, together with the propensity to colonize grape berry and the interesting enological traits possessed by the selected S. bacillaris strains allow the use of this yeast as biocontrol agent on vine and grape berries with possible positive effects on must fermentation, although the presence of S. cerevisiae is needed to complete the fermentation process. This work introduces

  5. Isolation and selection of local isolates of chitinolitic bacteria that potent to biocontrol of larva stadia of Aedes aegypti L.

    Directory of Open Access Journals (Sweden)

    SRI PUJIYANTO

    2008-01-01

    Full Text Available The dengue fever was the most dangerously epidemic in Indonesia, so it is important to prevent the population of Aedes aegypti L. The larva stages of Ae. aegypti had an exosceleton from chitin, so if its exosceleton had been degraded the larva would be died. Many bacteria had chitin degradation activities, and the bacteria had potential as a biocontrol for Ae. aegypti larva stages. This research was got many of the local chitinolitic bacteria that had been potential as a biocontrol for Ae. aegypti larva stages. The chitinolitic bacteria were isolated with selective agar medium. The sources of isolates were collected from Central Java and West Java water resources. Selected isolates was done to get the higher chitinolitic activity in mineral water medium. The isolate LMB1-5 was potential as a bioinsecticide for Ae. aegypti larva stages. This isolate decreased 86.7% of the larva population in 7 days.

  6. Ecological impacts of Phlebiopsis gigantea biocontrol treatment against Heterobasidion spp. as revealed by fungal community profiling and population analyses

    OpenAIRE

    Vainio, Eeva Johanna

    2008-01-01

    Wood decay fungi belonging to the species complex Heterobasidion annosum sensu lato are among the most common and economically important species causing root rot and stem decay in conifers of the northern temperate regions. New infections by these pathogens can be suppressed by tree stump treatments using chemical or biological control agents. In Finland, the corticiaceous fungus Phlebiopsis gigantea has been formulated into a commercial biocontrol agent called Rotstop (Verdera Ltd.). Th...

  7. Arbuscular mycorrhizal fungi for the biocontrol of plant-parasitic nematodes: a review of the mechanisms involved

    OpenAIRE

    Nele eSchouteden; Dirk eDe Waele; Bart ePanis; Christine M Vos

    2015-01-01

    Arbuscular mycorrhizal fungi (AMF) are obligate root symbionts that can protect their host plant against biotic stress factors such as plant parasitic nematode (PPN) infection. PPN consist of a wide range of species with different life styles that can cause major damage in many important crops worldwide. Various mechanisms have been proposed to play a role in the biocontrol effect of AMF against PPN. This review presents an overview of the different mechanisms that have been proposed, and dis...

  8. Arbuscular mycorrhizal fungi for the biocontrol of plant-parasitic nematodes: a review of the mechanisms involved

    OpenAIRE

    Schouteden, Nele; de Waele, Dirk; Panis, Bart; Christine M Vos

    2015-01-01

    Arbuscular mycorrhizal fungi (AMF) are obligate root symbionts that can protect their host plant against biotic stress factors such as plant-parasitic nematode (PPN) infection. PPN consist of a wide range of species with different life styles that can cause major damage in many important crops worldwide. Various mechanisms have been proposed to play a role in the biocontrol effect of AMF against PPN. This review presents an overview of the different mechanisms that have been proposed, and dis...

  9. Arbuscular Mycorrhizal Fungi for the Biocontrol of Plant-Parasitic Nematodes: A Review of the Mechanisms Involved.

    Science.gov (United States)

    Schouteden, Nele; De Waele, Dirk; Panis, Bart; Vos, Christine M

    2015-01-01

    Arbuscular mycorrhizal fungi (AMF) are obligate root symbionts that can protect their host plant against biotic stress factors such as plant-parasitic nematode (PPN) infection. PPN consist of a wide range of species with different life styles that can cause major damage in many important crops worldwide. Various mechanisms have been proposed to play a role in the biocontrol effect of AMF against PPN. This review presents an overview of the different mechanisms that have been proposed, and discusses into more detail the plausibility of their involvement in the biocontrol against PPN specifically. The proposed mechanisms include enhanced plant tolerance, direct competition for nutrients and space, induced systemic resistance (ISR) and altered rhizosphere interactions. Recent studies have emphasized the importance of ISR in biocontrol and are increasingly placing rhizosphere effects on the foreground as well, both of which will be the focal point of this review. Though AMF are not yet widely used in conventional agriculture, recent data help to develop a better insight into the modes of action, which will eventually lead toward future field applications of AMF against PPN. The scientific community has entered an exciting era that provides the tools to actually unravel the underlying molecular mechanisms, making this a timely opportunity for a review of our current knowledge and the challenges ahead. PMID:26635750

  10. Arbuscular mycorrhizal fungi for the biocontrol of plant-parasitic nematodes: a review of the mechanisms involved

    Directory of Open Access Journals (Sweden)

    Nele eSchouteden

    2015-11-01

    Full Text Available Arbuscular mycorrhizal fungi (AMF are obligate root symbionts that can protect their host plant against biotic stress factors such as plant parasitic nematode (PPN infection. PPN consist of a wide range of species with different life styles that can cause major damage in many important crops worldwide. Various mechanisms have been proposed to play a role in the biocontrol effect of AMF against PPN. This review presents an overview of the different mechanisms that have been proposed, and discusses into more detail the plausibility of their involvement in the biocontrol against PPN specifically. The proposed mechanisms include enhanced plant tolerance, direct competition for nutrients and space, induced systemic resistance (ISR and altered rhizosphere interactions. Recent studies have emphasized the importance of ISR in biocontrol and are increasingly placing rhizosphere effects on the foreground as well, both of which will be the focal point of this review. Though AMF are not yet widely used in conventional agriculture, recent data help to develop a better insight into the modes of action, which will eventually lead towards future field applications of AMF against PPN. The scientific community has entered an exciting era that provide the tools to actually unravel the underlying molecular mechanisms, making this a timely opportunity for a review of our current knowledge and the challenges ahead.

  11. Augmentative biocontrol in natural marine habitats: persistence, spread and non-target effects of the sea urchin Evechinus chloroticus.

    Science.gov (United States)

    Atalah, Javier; Hopkins, Grant A; Forrest, Barrie M

    2013-01-01

    Augmentative biocontrol aims to control established pest populations through enhancement of their indigenous enemies. To our knowledge, this approach has not been applied at an operational scale in natural marine habitats, in part because of the perceived risk of adverse non-target effects on native ecosystems. In this paper, we focus on the persistence, spread and non-target effects of the sea urchin Evechinus chloroticus when used as biocontrol agent to eradicate an invasive kelp from Fiordland, New Zealand. Rocky reef macrobenthic assemblages were monitored over 17 months in areas where the indigenous algal canopy was either removed or left intact prior to the translocation of a large number of urchins (>50 ind.·m(-2)). Urchin densities in treated areas significantly declined ∼9 months after transplant, and began spreading to adjacent sites. At the end of the 17-month study, densities had declined to ∼5 ind.·m(-2). Compared to controls, treatment sites showed persistent shifts from kelp forest to urchin barrens, which were accompanied by significant reductions in taxa richness. Although these non-target effects were pronounced, they were considered to be localised and reversible, and arguably outweigh the irreversible and more profound ecological impacts associated with the establishment of an invasive species in a region of high conservation value. Augmentative biocontrol, used in conjunction with traditional control methods, represents a promising tool for the integrated management of marine pests.

  12. Augmentative biocontrol in natural marine habitats: persistence, spread and non-target effects of the sea urchin Evechinus chloroticus.

    Directory of Open Access Journals (Sweden)

    Javier Atalah

    Full Text Available Augmentative biocontrol aims to control established pest populations through enhancement of their indigenous enemies. To our knowledge, this approach has not been applied at an operational scale in natural marine habitats, in part because of the perceived risk of adverse non-target effects on native ecosystems. In this paper, we focus on the persistence, spread and non-target effects of the sea urchin Evechinus chloroticus when used as biocontrol agent to eradicate an invasive kelp from Fiordland, New Zealand. Rocky reef macrobenthic assemblages were monitored over 17 months in areas where the indigenous algal canopy was either removed or left intact prior to the translocation of a large number of urchins (>50 ind.·m(-2. Urchin densities in treated areas significantly declined ∼9 months after transplant, and began spreading to adjacent sites. At the end of the 17-month study, densities had declined to ∼5 ind.·m(-2. Compared to controls, treatment sites showed persistent shifts from kelp forest to urchin barrens, which were accompanied by significant reductions in taxa richness. Although these non-target effects were pronounced, they were considered to be localised and reversible, and arguably outweigh the irreversible and more profound ecological impacts associated with the establishment of an invasive species in a region of high conservation value. Augmentative biocontrol, used in conjunction with traditional control methods, represents a promising tool for the integrated management of marine pests.

  13. Co-inoculation of aflatoxigenic and non-aflatoxigenic strains of Aspergillus flavus to study fungal invasion, colonization, and competition in maize kernels

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    Zuzana eHruska

    2014-03-01

    Full Text Available A currently utilized pre-harvest biocontrol method involves field inoculations with non-aflatoxigenic Aspergillus flavus strains, a tactic shown to strategically suppress native aflatoxin-producing strains and effectively decrease aflatoxin contamination in corn. The present in situ study focuses on tracking the invasion and colonization of an aflatoxigenic A. flavus strain (AF70, labeled with green fluorescent protein (GFP, in the presence of a non-aflatoxigenic A. flavus biocontrol strain (AF36 to better understand the competitive interaction between these two strains in seed tissue of corn (Zea mays. Corn kernels that had been co-inoculated with GFP-labeled AF70 and wild-type AF36 were cross-sectioned and observed under UV and blue light to determine the outcome of competition between these strains. After imaging, all kernels were analyzed for aflatoxin levels. There appeared to be a population difference between the co-inoculated AF70-GFP+AF36 and the individual AF70-GFP tests, both visually and with pixel count analysis. The GFP allowed us to observe that AF70-GFP inside the kernels was suppressed up to 82% when co-inoculated with AF36 indicating that AF36 inhibited progression of AF70-GFP. This was in agreement with images taken of whole kernels where AF36 exhibited a more robust external growth compared to AF70-GFP. The suppressed growth of AF70-GFP was reflected in a corresponding (up to 73% suppression in aflatoxin levels. Our results indicate that the decrease in aflatoxin production correlated with population depression of the aflatoxigenic fungus by the biocontrol strain supporting the theory of competitive exclusion through robust propagation and fast colonization by the non-aflatoxigenic fungus.

  14. IMPROVMENT BIOCONTROL OF DAMPING-OFF AND ROOT ROT/WILT OF FABA BEAN BY SALICYLIC ACID AND HYDROGEN PEROXIDE

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    Montaser F. Abdel-Monaim

    2013-04-01

    Full Text Available Rhizoctonia solani, Fusarium solani, F. oxysporum and Macrophomina phaseolina were found to be associated with root rot and wilt symptoms of faba bean plants collected from different fields in New Valley governorate, Egypt. All the obtained isolates were able to attack faba bean plants (cv. Giza 40 causing damping-off and root rot/wilt diseases. R. solani isolates 2 and 5, F. solani isolate 8, F. oxysporum isolate 12 and M. phaseolina isolate 14 were the more virulent ones in the pathogenicity tests. Biocontrol agents (Trichoderma viride, Bacillus megaterium and chemical inducers (salicylic acid and hydrogen peroxide individually or in combination were examined for biological control of damping-off and root rot/wilt and growth promotion of faba bean plants in vitro and in vivo. Both antagonistic biocontrol agents and chemical inducers either individually or in combination inhibited growth of the tested pathogenic fungi.Biocontrol agents combined with chemical inducers recorded the highest inhibited growth especially in case SA + T. viride and SA+ B. megaterium. Under green house and field conditions, all treatments significantly reduced damping-off and root rot/ wilt severity and increased survival of plants. Also, these treatments increased fresh and dry weights of the survived plants in pots compared with control.  The combination of biocontrol agents and chemical inducers were more effective than using them individually and SA+ T. viride was the best treatment in this respect. Under field conditions, all these treatments significantly increased growth parameters (plant height and No. of branches plant-1 and yield components (No. of pods and seedsplant-1, weight of 100 seeds and total yield feddan-1 and protein content in both seasons (2010-2011 and 2011-2012. Faba bean seeds soaked in SA+ T. viride and SA+ B. megaterium were recorded the highest growth parameters and yield components. Generally, the combination of biocontrol agents and

  15. Suppression of the biocontrol agent trichoderma harzianum by mycelium of the arbuscular mycorrhizal fungus glomus intraradices in root-free soil

    Science.gov (United States)

    Green; Larsen; Olsson; Jensen; Jakobsen

    1999-04-01

    Trichoderma harzianum is an effective biocontrol agent against several fungal soilborne plant pathogens. However, possible adverse effects of this fungus on arbuscular mycorrhizal fungi might be a drawback in its use in plant protection. The objective of the present work was to examine the interaction between Glomus intraradices and T. harzianum in soil. The use of a compartmented growth system with root-free soil compartments enabled us to study fungal interactions without the interfering effects of roots. Growth of the fungi was monitored by measuring hyphal length and population densities, while specific fatty acid signatures were used as indicators of living fungal biomass. Hyphal 33P transport and beta-glucuronidase (GUS) activity were used to monitor activity of G. intraradices and a GUS-transformed strain of T. harzianum, respectively. As growth and metabolism of T. harzianum are requirements for antagonism, the impact of wheat bran, added as an organic nutrient source for T. harzianum, was investigated. The presence of T. harzianum in root-free soil reduced root colonization by G. intraradices. The external hyphal length density of G. intraradices was reduced by the presence of T. harzianum in combination with wheat bran, but the living hyphal biomass, measured as the content of a membrane fatty acid, was not reduced. Hyphal 33P transport by G. intraradices also was not affected by T. harzianum. This suggests that T. harzianum exploited the dead mycelium but not the living biomass of G. intraradices. The presence of external mycelium of G. intraradices suppressed T. harzianum population development and GUS activity. Stimulation of the hyphal biomass of G. intraradices by organic amendment suggests that nutrient competition is a likely means of interaction. In conclusion, it seemed that growth of and phosphorus uptake by the external mycelium of G. intraradices were not affected by the antagonistic fungus T. harzianum; in contrast, T. harzianum was adversely

  16. DNA-based methodologies for the quantification of live and dead cells in formulated biocontrol products based on Pantoea agglomerans CPA-2.

    Science.gov (United States)

    Soto-Muñoz, Lourdes; Torres, Rosario; Usall, Josep; Viñas, Inmaculada; Solsona, Cristina; Teixidó, Neus

    2015-10-01

    Pantoea agglomerans strain CPA-2 is an effective biocontrol agent (BCA) against the major postharvest pathogens present on pome and citrus fruits. Dehydration, such as freeze-drying, spray-drying and fluidized bed drying is one of the best ways to formulate BCAs. In this work, the survival of CPA-2 cells after formulation was determined by dilution plating and molecular methods as qPCR alone and combined with a sample pretreatment with a propidium monoazide dye (PMA-qPCR) and they were used to calculate treatment concentrations in efficacy trials on postharvest oranges. Furthermore, no significant differences in CPA-2 survival were observed as determined by dilution plating and PMA-qPCR after both the freeze drying and fluidized bed drying processes; however, an interesting significant difference was observed in the spray dried product comparing all quantitative methods. A difference of 0.48 and 2.17 log10 CFU or cells g/dw was observed among PMA-qPCR with qPCR and dilution plating, respectively. According to our study, dilution plating was shown to be an unreliable tool for monitoring the survival of CPA-2 after spray drying. In contrast, the combination of PMA and qPCR enabled a quick and unequivocal methodology to enumerate viable and VBNC CPA-2 cells under stress-dried conditions. Efficacy trials showed that, after 3 days, spray drying formulation rehydrated with 10% non-fat skimmed milk (NFSM) was as effective as fresh cells to control Penicillium digitatum in oranges. PMID:26114590

  17. Evaluation of potential antagonistism in yeasts, seeking biocontrol of spoilage by Penicillium expansumAvaliação do potencial antagônico de leveduras, visando biocontrole de deterioração por Penicillium expansum

    Directory of Open Access Journals (Sweden)

    Kei-ichi Harada

    2011-12-01

    Full Text Available Considerable losses during apple fruit storage occur due to microbiological diseases, mainly caused by Penicillium expansum, which in addition to fruit pulp deterioration produces patulin, a mycotoxin with carcinogenic and teratogenic activity. Biological control of post-harvest disease by antagonist yeasts focused on killer toxins is an appreciable alternative to the chemical fungicides, due to the low possibility of toxic residues demonstrated during fermentative processes. Twenty out of 44 yeasts (16 isolated from fruits, 10 from corn silage and 18 from laboratory anthill, showed antagonism against spores of P. expansum. The assay in solid medium pointed the strongest nutrient competition antagonism by D. hansenii strain C1 (31 mm inhibition diameter, while D. hansenii strain C7 (15 mm showed higher antibiosis and parasitism pattern. In the following step the extracellular activity was tested performing the assay with culture supernatant in Yeast Medium agar, where C. guilliermondii P3 was more effective against conidia germination (inhibition rate of 58.15% while P. ohmeri showed better inhibition on micelial growth (66.17%. The antibiosis showed by both yeasts could suggest probable mechanism associated with killer phenomenon, once both strains were killer positive against sensitive reference strains (S. cerevisiae NCYC 1006 and P. kluyveri CAY-15. In order to enhance the production of antifungal substance, these yeasts were cultivated with P. expansum, but the difference between culture supernatant obtained from yeasts cultivated alone and with mould was not significant (P > 0.05. The results demonstrated that the yeasts application constitute a promising tool, enhancing the biological control of P. expansum in post-harvest diseases of apple fruit.As perdas consideráveis no armazenamento de maçãs decorrem principalmente de desordens microbiológicas, causadas por Penicillium expansum, que além de colonizar o fruto e causar dano

  18. The toolbox of Trichoderma spp. in the biocontrol of Botrytis cinerea disease.

    Science.gov (United States)

    Vos, Christine M F; De Cremer, Kaat; Cammue, Bruno P A; De Coninck, Barbara

    2015-05-01

    Botrytis cinerea is a necrotrophic fungal pathogen causing disease in many plant species, leading to economically important crop losses. So far, fungicides have been widely used to control this pathogen. However, in addition to their detrimental effects on the environment and potential risks for human health, increasing fungicide resistance has been observed in the B. cinerea population. Biological control, that is the application of microbial organisms to reduce disease, has gained importance as an alternative or complementary approach to fungicides. In this respect, the genus Trichoderma constitutes a promising pool of organisms with potential for B. cinerea control. In the first part of this article, we review the specific mechanisms involved in the direct interaction between the two fungi, including mycoparasitism, the production of antimicrobial compounds and enzymes (collectively called antagonism), and competition for nutrients and space. In addition, biocontrol has also been observed when Trichoderma is physically separated from the pathogen, thus implying an indirect systemic plant defence response. Therefore, in the second part, we describe the consecutive steps leading to induced systemic resistance (ISR), starting with the initial Trichoderma-plant interaction and followed by the activation of downstream signal transduction pathways and, ultimately, the defence response resulting in ISR (ISR-prime phase). Finally, we discuss the ISR-boost phase, representing the effect of ISR priming by Trichoderma spp. on plant responses after additional challenge with B. cinerea.

  19. The advantages of mosquito biocontrol by stocking edible fish in rice paddies.

    Science.gov (United States)

    Wu, N; Liao, G H; Li, D F; Luo, Y L; Zhong, G M

    1991-09-01

    Edible fish stocked in rice fields at a density of 600-800 fry per mu (1 mu = 1/15 hectare) for 150-170 days may act as an effective mosquito biocontrol agent. Common carp (Cyprinus carpio), grass carp Ctenopharyngodon idella) and Tilopia spp. killed late stage larvae and pupae of Anopheles sinensis and Culex tritaeniorrhyncus in laboratory and field trials. Stocking of fish in experimental rice fields decreased larval numbers significantly in comparison with control areas. Expansion of fish stocking in rice fields on a large scale over several years correlated with a marked decrease in malaria transmission. The addition of fish to the rice fields also resulted in increased yields. A ditch-ridge system of field arrangements is described for optimization of fish handling. Preliminary cost-benefit analysis indicates that this approach to mosquito control conveys considerable economic advantage and thus provides incentive to the community to participate in vector control programs. Farmers' experience in Guangxi over a number of years indicates that the use of edible fish for this purpose can be carried on a large, commercially viable scale.

  20. Chitin enhances biocontrol of Rhodotorula mucilaginosa to postharvest decay of peaches.

    Science.gov (United States)

    Zhang, Hongyin; Yang, Qiya; Ge, Lingling; Zhang, Guochao; Zhang, Xiaoli; Zhang, Xiaoyun

    2016-07-01

    Biological control using microbial antagonists is a promising alternative approach to synthetic fungicides. However, effective biological control requires enhancing the consistency and efficacy of the antagonists used to control postharvest diseases. This study investigated the effect of chitin on the biocontrol efficacy of Rhodotorula mucilaginosa against blue mold and Rhizopus decay of peaches and on the protein expression profiles of R. mucilaginosa. The antagonistic activity of R. mucilaginosa harvested from the nutrient yeast dextrose broth (NYDB) with 0.5% chitin added was significantly improved compared with culture in NYDB without chitin. The R. mucilaginosa population cultured in chitin-supplement NYDB and nutrient yeast chitin borth (NYCB) harvested from peach wounds was more than that of R. mucilaginosa cultured in NYDB without chitin throughout the storage period except at 1 d. The protein expression profiles findings revealed that there were several differentially expressed proteins of R. mucilaginosa in the 0.5% chitin-supplemented NYDB and NYCB compared with that of R. mucilaginosa in NYDB. Most of these were cellular proteomes relating to the primary metabolic reactions such as glycoside hydrolases, phosphoribosyl pyrophosphate, and NADH dehydrogenases. Some proteins were also related to signal transmission and stress response. PMID:27064085

  1. Purification of an antifungal endochitinase from a potential biocontrol Agent Streptomyces griseus.

    Science.gov (United States)

    Rabeeth, M; Anitha, A; Srikanth, Geetha

    2011-08-15

    Streptomyces griseus (MTCC 9723) is a chitinolytic bacterium isolated from prawn cultivated pond soil of Peddapuram Village; East Godavari District was studied in detailed. Chitinase (EC 3.2.1.14) was extracted from the culture filtrate of Streptomyces griseus and purified by ammonium sulfate precipitation, DEAE-cellulose ionexchange chromatography, Sephadex G-100 and Sephadex G-200 gel filtration chromatography. The molecular mass of the purified chitinase was estimated to be 34, 32 kDa by SDS gel electrophoresis and confirmed by activity staining with Calcofluor White M2R. Chitinase was optimally active at pH of 6.0 and at 40 degrees C. The enzyme was stable from pH 5-9 and up to 20-50 degrees C. The chitinase exhibited Km and Vmax values of 400 mg and 180 IU mL(-1) for colloidal chitin. Among the metals and inhibitors that were tested, the Hg+, Hg2+ and P-chloromercuribenzoic acid completely inhibited the chitinase activity at 1 mM concentration. The purified chitinase showed high activity on colloidal chitin, chitobiose, and chitooligosaccharide. An in vitro assay proved that the crude chitinase, actively growing cells of S. griseus having antifungal activity against all studied fungal pathogen. This result implies that characteristics of S. griseus producing endochitinase made them suitable for biotechnological purpose such as for degradation of chitin containing waste and it might be a promising biocontrol agent for plant pathogens. PMID:22545353

  2. Biocontrol mechanisms of Trichoderma harzianum against soybean charcoal rot caused by Macrophomina phaseolina

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    Khaledi Nima

    2016-01-01

    Full Text Available Throughout the world, charcoal rot, caused by Macrophomina phaseolina, is one of the most destructive and widespread diseases of crop plants such as soybean. In this study, the biological control capability of 11 Trichoderma spp. isolates against M. phaseolina was investigated using screening tests. Among all the tested Trichoderma spp. isolates, inhibition varied from 20.22 to 58.67% in dual culture tests. Dual culture, volatile and non-volatile tests revealed that two isolates of Trichoderma harzianum (including the isolates T7 and T14 best inhibited the growth of M. phaseolina in vitro. Therefore, these isolates were selected for biocontrol of M. phaseolina in vivo. The results of greenhouse experiments revealed that disease severity in the seed treatment with T. harzianum isolates was significantly lower than that of the soil treatment. In most of the cases, though, soil treatment with T. harzianum resulted in higher plant growth parameters, such as root and shoot weight. The effects of T. harzianum isolates on the activity of peroxidase enzyme and phenolic contents of the soybean root in the presence and absence of M. phaseolina were determined in greenhouse conditions. Our results suggested that a part of the inhibitory effect of T. harzianum isolates on soybean charcoal rot might be related to the indirect influence on M. phaseolina. Plant defense responses were activated as an elicitor in addition to the direct effect on the pathogen growth.

  3. Molecular characterization of alkaline protease of Bacillus amyloliquefaciens SP1 involved in biocontrol of Fusarium oxysporum.

    Science.gov (United States)

    Guleria, Shiwani; Walia, Abhishek; Chauhan, Anjali; Shirkot, C K

    2016-09-01

    An alkaline protease gene was amplified from genomic DNA of Bacillus amyloliquefaciens SP1 which was involved in effective biocontrol of Fusarium oxysporum. We investigated the antagonistic capacity of protease of B. amyloliquifaciens SP1, under in vitro conditions. The 5.62 fold purified enzyme with specific activity of 607.69U/mg reported 24.14% growth inhibition of F. oxysporum. However, no antagonistic activity was found after addition of protease inhibitor i.e. PMSF (15mM) to purified enzyme. An 1149bp nucleotide sequence of protease gene encoded 382 amino acids of 43kDa and calculated isoelectric point of 9.29. Analysis of deduced amino acid sequence revealed high homology (86%) with subtilisin E of Bacillus subtilis. The B. amyloliquefaciens SP1 protease gene was expressed in Escherichiax coli BL21. The expressed protease was secreted into culture medium by E. coli and exhibited optimum activity at pH8.0 and 60°C. The most reliable three dimensional structure of alkaline protease was determined using Phyre 2 server which was validated on the basis of Ramachandran plot and ERRAT value. The expression and structure prediction of the enzyme offers potential value for commercial application in agriculture and industry. PMID:27294522

  4. Ecology of Anti-Biofilm Agents II: Bacteriophage Exploitation and Biocontrol of Biofilm Bacteria

    Directory of Open Access Journals (Sweden)

    Stephen T. Abedon

    2015-09-01

    Full Text Available Bacteriophages are the viruses of bacteria. In the guise of phage therapy they have been used for decades to successfully treat what are probable biofilm-containing chronic bacterial infections. More recently, phage treatment or biocontrol of biofilm bacteria has been brought back to the laboratory for more rigorous assessment as well as towards the use of phages to combat environmental biofilms, ones other than those directly associated with bacterial infections. Considered in a companion article is the inherent ecological utility of bacteriophages versus antibiotics as anti-biofilm agents. Discussed here is a model for phage ecological interaction with bacteria as they may occur across biofilm-containing ecosystems. Specifically, to the extent that individual bacterial types are not highly abundant within biofilm-containing environments, then phage exploitation of those bacteria may represent a “Feast-or-famine” existence in which infection of highly localized concentrations of phage-sensitive bacteria alternate with treacherous searches by the resulting phage progeny virions for new concentrations of phage-sensitive bacteria to infect. An updated synopsis of the literature concerning laboratory testing of phage use to combat bacterial biofilms is then provided along with tips on how “Ecologically” such phage-mediated biofilm control can be modified to more reliably achieve anti-biofilm efficacy.

  5. The toolbox of Trichoderma spp. in the biocontrol of Botrytis cinerea disease.

    Science.gov (United States)

    Vos, Christine M F; De Cremer, Kaat; Cammue, Bruno P A; De Coninck, Barbara

    2015-05-01

    Botrytis cinerea is a necrotrophic fungal pathogen causing disease in many plant species, leading to economically important crop losses. So far, fungicides have been widely used to control this pathogen. However, in addition to their detrimental effects on the environment and potential risks for human health, increasing fungicide resistance has been observed in the B. cinerea population. Biological control, that is the application of microbial organisms to reduce disease, has gained importance as an alternative or complementary approach to fungicides. In this respect, the genus Trichoderma constitutes a promising pool of organisms with potential for B. cinerea control. In the first part of this article, we review the specific mechanisms involved in the direct interaction between the two fungi, including mycoparasitism, the production of antimicrobial compounds and enzymes (collectively called antagonism), and competition for nutrients and space. In addition, biocontrol has also been observed when Trichoderma is physically separated from the pathogen, thus implying an indirect systemic plant defence response. Therefore, in the second part, we describe the consecutive steps leading to induced systemic resistance (ISR), starting with the initial Trichoderma-plant interaction and followed by the activation of downstream signal transduction pathways and, ultimately, the defence response resulting in ISR (ISR-prime phase). Finally, we discuss the ISR-boost phase, representing the effect of ISR priming by Trichoderma spp. on plant responses after additional challenge with B. cinerea. PMID:25171761

  6. Biocontrol of seed pathogens and growth promotion of common bean seedlings by Trichoderma harzianum

    Directory of Open Access Journals (Sweden)

    Daniel Diego Costa Carvalho

    2011-08-01

    Full Text Available The objective of this work was to evaluate isolates of Trichoderma harzianum regarding biocontrol of common bean seed-borne pathogens, plant growth promotion, and rhizosphere competence. Five isolates of T. harzianum were evaluated and compared with commercial isolate (Ecotrich, Carboxin+Thiram, and an absolute control. Bean seeds of the cultivar Jalo Precoce, contaminated with Aspergillus, Cladosporium, and Sclerotinia sclerotiorum, were microbiolized with antagonists, and seed health tests were carried out. Isolates were evaluated on autoclaved substrate and in field conditions. Ten days after sowing (DAS, plant length was measured. To test rhizosphere competence, isolates were applied in boxes containing autoclaved washed sand, and root colonization was evaluated at 10 DAS, using five plants per box. The most effective isolates in the seed health tests were: CEN287 and CEN289 to control Aspergillus; the commercial isolate to control Cladosporium; and CEN287 and CEN316 to control S. sclerotiorum. Isolates CEN289 and CEN290 promoted bean growth in greenhouse and field. Seed treatment with T. harzianum reduces the incidence of Aspergillus, Cladosporium, and S. sclerotiorum in 'Jalo Precoce' common bean seeds.

  7. Plot- and landscape-level changes in climate and vegetation following defoliation of exotic saltcedar (Tamarix sp.) from the biocontrol agent Diorhabda carinulata along a stream in the Mojave Desert (USA)

    Science.gov (United States)

    Bateman, H.L.; Nagler, P.L.; Glenn, E.P.

    2013-01-01

    The biocontrol agent, northern tamarisk beetle (Diorhabda carinulata), has been used to defoliate non-native saltcedar (Tamarix spp.) in USA western riparian systems since 2001. Biocontrol has the potential to impact biotic communities and climatic conditions in affected riparian areas. To determine the relationships between biocontrol establishment and effects on vegetation and climate at the plot and landscape scales, we measured temperature, relative humidity, foliage canopy, solar radiation, and used satellite imagery to assess saltcedar defoliation and evapotranspiration (ET) along the Virgin River in the Mojave Desert. Following defoliation solar radiation increased, daily humidity decreased, and maximum daily temperatures tended to increase. MODIS and Landsat satellite imagery showed defoliation was widespread, resulting in reductions in ET and vegetation indices. Because biocontrol beetles are spreading into new saltcedar habitats on arid western rivers, and the eventual equilibrium between beetles and saltcedar is unknown, it is necessary to monitor trends for ecosystem functions and higher trophic-level responses in habitats impacted by biocontrol.

  8. Evaluation of yeasts obtained from Antarctic soil samples as biocontrol agents for the management of postharvest diseases of apple (Malus × domestica).

    Science.gov (United States)

    Vero, Silvana; Garmendia, Gabriela; González, M Belén; Bentancur, Oscar; Wisniewski, Michael

    2013-03-01

    Psychrotrophic yeasts were isolated from Antarctic soils, selected based on their ability to grow in apple juice at low temperatures, and were evaluated as potential biocontrol agents for the management of postharvest diseases of apple during cold storage. Among the species recovered, an isolate of Leucosporidium scottii, designated At17, was identified as a good biocontrol agent for blue and gray mold of two apple cultivars. The selected isolate produced soluble and volatile antifungal substances that were inhibitory to apple pathogens. Siderophore production was also demonstrated, but it did not appear to play a role in pathogen inhibition. The selected yeast had the capacity to form a biofilm when grown in apple juice, which is considered an important attribute of postharvest antagonists to successfully colonize wounds and intact fruit surfaces. At17 was resistant to commonly used postharvest fungicides, so application of a combination of low-dose fungicide along with the biocontrol agent could be used as an integrated management practice.

  9. Screening of Pseudomonas sp. Isolated from Rhizosphere of Soybean Plant as Plant Growth Promoter and Biocontrol Agent

    Directory of Open Access Journals (Sweden)

    Aris T. Wahyudi

    2011-01-01

    Full Text Available Problem statement: Pseudomonas spesies are one of the rihizobacterial group that have an important role in plant growth promoter and plant health. To prepare them as inoculants, they must have a range of characters as growth promoter such as Indole Acetic Acid (IAA producers which can promote the growth of plants and solubilize phosphates. In addition, they must also have the various characters that act as biocontrol agents such as siderofor, chitinase and anti-fungal compound producers. Approach: Pseudomonas sp isolated from soybeans rhizospere and identified based on physiological reactions and 16S rRNA gene sequences. Various tests for the determination of the growth promoter were based on IAA production, phosphate solubilization and growth promoter of length of root and stems and number of lateral roots of soybean sprouts. Test of siderophore, chitinase, as well as anti anti-fungal compounds productions to inhibit the growth of Fusarium oxysporum, Rhizoctonia solani and Sclerotium rolfsii, were used as a biocontrol agent determination. Hypersensitivity test was used to screen for Pseudomonas sp classified as non-pathogenic rhizobacteria. Results: Fourteen isolates identified as a non-pathogenic Pseudomonas sp that produced IAA and Promoted enhancement of root length, shoot length, or number of lateral root. Among those 14 isolates, 8 isolates showed phosphate solubilizing activity, 12 isolates capable of producing siderophore and six isolates were observed to have chitinolytic activity. Only three isolates were able to inhibit the growth of Fusarium oxysporum in high level. While one and two isolates inhibited Sclerotium rolfsii and Rhizoctonia solani in high level, respectively. Conclusion: On the basis of excellent growth promoter and biocontrol activities, we recommended 5 isolates of Pseudomonas sp which were Crb-3, Crb-16, Crb-17, Crb-44 and Crb-94 as potential isolates of Pseudomonas sp that could be applied as

  10. Induction of banana Fusarium Wilt disease with biocontrol bacteria and pathogen by inoculation%接种生防菌和病原菌对香蕉抗枯萎病的诱导

    Institute of Scientific and Technical Information of China (English)

    薛玉潇; 贾慧升; 王国芬; 刘磊; 黄俊生

    2012-01-01

    Banana vitroplants of 4--5 leaves were inoculated in different ways with two biocontrol bacteria Bacillus subtilis strains A5-6 and C10-1 and the banana Fusarium wilt pathogen (Foc4) to observe their changes of polyphenol oxidase ( PPO), peroxidase (POD) and phenylalanine ammonialyase (PAL) activities in leaves, which are related to plant tolerance to the disease. The PPO, POD and PAL activities in the leaves of the banana plants were determined by using the physiological and biochemical methods. The banana plants inoculated with the strains A5 -6 and C10-1 separately showed higher leaf PPO, POD and PAL activities than those inoculated with the sterile water control, and the banana plants inoculated with mixture of the biocontrol bacteria and Foc4 pathogen had higher leaf PPO, POD and PAL activities than those inoculated with Foc4 alone.%使用2株生防枯草芽孢杆菌A5-6,C10-1和香蕉枯萎病病原菌(Foc4),以不同的接种方式对4~5叶香蕉组培苗进行根部接种,以苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、多酚氧化酶(PPO)这3种与植物抗病性密切相关的酶活性变化为指标,研究植物产生抗性的时间和强度的变化趋势。结果表明,经过生防菌A5-6和C10-1诱导处理的叶片PAL,POD,PPO的活性均高于对照(CK),生防菌和病原菌混合接种处理的POD和PPO酶活峰值高于生防菌和病原菌单独接种的处理。

  11. New solid-state fermentation chamber for bulk production of aerial conidia of fungal biocontrol agents on rice.

    Science.gov (United States)

    Ye, S D; Ying, S H; Chen, C; Feng, M G

    2006-06-01

    A novel solid-state fermentation apparatus, namely an upright multi-tray conidiation chamber, was developed to facilitate the production of aerial conidia of fungal biocontrol agents, such as Beauveria bassiana. The chamber with 25 bottom-meshed metal trays had a capacity of > or =50 kg rice with each tray holding > or =2 kg. In repeated trials, a mean yield of 2.4 (1.8-2.7) x 10(12) conidia kg(-1) rice was harvested from the 7-day cultures of B. bassiana in a fully loaded chamber. The new apparatus has a high potential for bulk production of fungal conidia. PMID:16786244

  12. Shallot Aphids, Myzus ascalonicus, in Strawberry: Biocontrol Potential of Three Predators and Three Parasitoids

    DEFF Research Database (Denmark)

    Enkegaard, Annie; Sigsgaard, Lene; Kristensen, Kristian

    2013-01-01

    or sweet pepper leaves. The aphid species and the plant species did not, however, have a significant influence on the number of females laying eggs, the average number of eggs laid during the first day being 6.37±1.28 per female. Adult lady beetles had a significant preference for odor from controls...... without plants over odors from uninfested strawberry or pepper plants, but they showed no preference for either of the plant species, whether infested with aphids or not. The predation capacity of A. bipunctata on shallot aphids holds promise for its use in inundative biocontrol, and the results on egg...

  13. Evaluation of the enzymatic activity of Trichoderma inhamatum (BOL-12QD as possible biocontroller

    Directory of Open Access Journals (Sweden)

    García-Espejo Cielo Noemí

    2016-02-01

    Full Text Available It is known that Trichoderma spp. acts as a natural biocontroller of pathogen fungi, is for this reason, that this research studies the potential of its hydrolytic enzyme activity. In this article, first we determined that the speed of growth of Trichoderma inhamatum cepa BOL-12QD is 9 hours. Later, we proposed a simple and sensitive method based in the use of basal media (BM with coloidal chitin as the only carbon resource and supplemented with bromocresol purple for the qualitative determination of chitinase activi-ty. On the other hand, it was determined the celullolytic and proteolytic activities of Trichoderma in-hamatum cepa BOL-12QD and it was observed that agitation, type and concentration of sustrate are determinant factors in enzymatic production. Then, we evaluated the cellulolytic activity of Trichoderma inhamatum cepa BOL-12QD in agitation and stationary using carboxymethylcellulose (CMC as sustrate, finding that using a 2% of sustrate the highest activity is registered at 8 days of incubation in agitation with a value of 99.23 IU/L. In relation to the results at stationary the optimal value is at the fourth day with a value of 92.76 IU/L. The protease activity it was determined taking in consideration variables of agitation and stationary, using different types and concentration of sustrate at 2%, 4% y 6% (w/v of meat extract, 1%, 3%, 5% (w/v of jelly and 1%, 2%, 4% (w/v of casein. The highest protease activity was obtained at the end of the sev-enth day with an enzymatic activity of 3075.45 IU/L at stationary using a concentration of 6% (w/v of meat extract, and using jelly at 3% (w/v at stationary it was found an activity of 568.36 IU/L on the tenth day and in agitation a value of 547.27 IU/L was reached on the twelveth day, while using casein at 1% (w/v at stationary an activity of 407.06 IU/L is reached in the fifth day, and in agitation at 4% (w/v of casein a value of 547.27 IU/L is obtained on the twelfth day, while using

  14. Biocontrol efficacy and plant growth promoting activity of Bacillus altitudinis isolated from Darjeeling hills, India.

    Science.gov (United States)

    Sunar, Kiran; Dey, Pannalal; Chakraborty, Usha; Chakraborty, Bishwanath

    2015-01-01

    A total of 18 bacterial isolates were obtained from the rhizosphere of Sechium edule growing in the lower foothills of Darjeeling, India. The bacterial isolates were tested for PGPR traits in vitro such as phosphate solubilization, HCN, siderophore, IAA, chitinase, protease production as well as inhibition of pthytopathogens. Of all the bacterial isolates, one bacterium designated as BRHS/S-73 was found to possess all the tested characters which was identified on the basis of 16S rRNA gene sequence analysis as Bacillus altitudinis and was selected for in vivo studies. A significant improvement in growth measured in terms of increase in root length, shoot length, and increase in root and shoot biomass was observed when seeds of Vigna radiata, Cicer arietinum, and Glycine max were bacterized prior to sowing in field condition. Besides, the bacterium could also solubilize soil phosphate. Apart form growth promotion, root rot disease of Vigna radiata caused by Thanatephorus cucumeris was also significantly reduced by 74% when the bacterium was applied to the rhizosphere prior to pathogen challenge. The biocontrol efficacy of the bacterium was found to be 66.6% even after 30 days of pathogen inoculation. Activities of key defense related enzymes such as phenylalanine ammonia lyase, peroxidase, β-1,3-glucanase, and chitinase in both roots and leaves of treated plants were also enhanced. Results clearly suggest that B. altitudinis (BRHS/S-73) is a potential PGPR which can be used as efficient microorganism for enhancement of plant growth and suppression of fungal disease. PMID:23996212

  15. Biocontrol and Rapid Detection of Food-Borne Pathogens Using Bacteriophages and Endolysins.

    Science.gov (United States)

    Bai, Jaewoo; Kim, You-Tae; Ryu, Sangryeol; Lee, Ju-Hoon

    2016-01-01

    Bacteriophages have been suggested as natural food preservatives as well as rapid detection materials for food-borne pathogens in various foods. Since Listeria monocytogenes-targeting phage cocktail (ListShield) was approved for applications in foods, numerous phages have been screened and experimentally characterized for phage applications in foods. A single phage and phage cocktail treatments to various foods contaminated with food-borne pathogens including E. coli O157:H7, Salmonella enterica, Campylobacter jejuni, Listeria monocytogenes, Staphylococcus aureus, Cronobacter sakazakii, and Vibrio spp. revealed that they have great potential to control various food-borne pathogens and may be alternative for conventional food preservatives. In addition, phage-derived endolysins with high host specificity and host lysis activities may be preferred to food applications rather than phages. For rapid detection of food-borne pathogens, cell-wall binding domains (CBDs) from endolysins have been suggested due to their high host-specific binding. Fluorescence-tagged CBDs have been successfully evaluated and suggested to be alternative materials of expensive antibodies for various detection applications. Most recently, reporter phage systems have been developed and tested to confirm their usability and accuracy for specific detection. These systems revealed some advantages like rapid detection of only viable pathogenic cells without interference by food components in a very short reaction time, suggesting that these systems may be suitable for monitoring of pathogens in foods. Consequently, phage is the next-generation biocontrol agent as well as rapid detection tool to confirm and even identify the food-borne pathogens present in various foods.

  16. Biocontrol and Rapid Detection of Food-borne Pathogens Using Bacteriophages and Endolysins

    Directory of Open Access Journals (Sweden)

    Jaewoo eBai

    2016-04-01

    Full Text Available Bacteriophages have been suggested as natural food preservatives as well as rapid detection materials for food-borne pathogens in various foods. Since Listeria monocytogenes-targeting phage cocktail (ListShield was approved for applications in foods, numerous phages have been screened and experimentally characterized for phage applications in foods. A single phage and phage cocktail treatments to various foods contaminated with food-borne pathogens including E. coli O157:H7, Salmonella enterica, Campylobacter jejuni, Listeria monocytogenes, Staphylococcus aureus, Cronobacter sakazakii, and Vibrio spp. revealed that they have great potential to control various food-borne pathogens and may be alternative for conventional food preservatives. In addition, phage-derived endolysins with high host specificity and host lysis activities may be preferred to food applications rather than phages. For rapid detection of food-borne pathogens, cell-wall binding domains (CBDs from endolysins have been suggested due to their high host-specific binding. Fluorescence-tagged CBDs have been successfully evaluated and suggested to be alternative materials of expensive antibodies for various detection applications. Most recently, reporter phage systems have been developed and tested to confirm their usability and accuracy for specific detection. These systems revealed some advantages like rapid detection of only viable pathogenic cells without interference by food components in a very short reaction time, suggesting that these systems may be suitable for monitoring of pathogens in foods. Consequently, phage is the next-generation biocontrol agent as well as rapid detection tool to confirm and even identify the food-borne pathogens present in various foods.

  17. Amblyseius swirskii: what made this predatory mite such a successful biocontrol agent?

    Science.gov (United States)

    Calvo, F Javier; Knapp, Markus; van Houten, Yvonne M; Hoogerbrugge, Hans; Belda, José E

    2015-04-01

    The predatory mite Amblyseius swirskii quickly became one of the most successful biocontrol agents in protected cultivation after its introduction into the market in 2005 and is now released in more than 50 countries. There are several key factors contributing to this success: (1) it can control several major pests including the western flower thrips, Frankliniella occidentalis, the whiteflies Bemisia tabaci and Trialeurodes vaporariorum and the broad mite, Polyphagotarsonemus latus, simultaneously in vegetables and ornamental crops; (2) it can develop and reproduce feeding on non-prey food sources such as pollen, which allows populations of the predator to build up on plants before the pests are present and to persist in the crop during periods when prey is scarce or absent; and (3) it can be easily reared on factitious prey, which allows economic mass production. However, despite the fact that A. swirskii provides growers with a robust control method, external demands were initially a key factor in promoting the use of this predator, particularly in Spain. In 2006, when exports of fresh vegetables from Spain were stopped due to the presence of pesticide residues, growers were forced to look for alternatives to chemical control. This resulted in the massive adoption of biological control-based integrated pest management programmes based on the use of A. swirskii in sweet pepper. Biological control increased from 5 % in 2005, 1 year before A. swirskii was commercially released, to almost 100 % of a total 6,000 ha of protected sweet pepper in Spain within 3 years. Later, it was demonstrated that A. swirskii was equally effective in other crops and countries, resulting in extensive worldwide use of A. swirskii in greenhouses.

  18. What magnitude are observed non-target impacts from weed biocontrol?

    Science.gov (United States)

    Suckling, David Maxwell; Sforza, René François Henri

    2014-01-01

    A systematic review focused by plant on non-target impacts from agents deliberately introduced for the biological control of weeds found significant non-target impacts to be rare. The magnitude of direct impact of 43 biocontrol agents on 140 non-target plants was retrospectively categorized using a risk management framework for ecological impacts of invasive species (minimal, minor, moderate, major, massive). The vast majority of agents introduced for classical biological control of weeds (>99% of 512 agents released) have had no known significant adverse effects on non-target plants thus far; major effects suppressing non-target plant populations could be expected to be detectable. Most direct non-target impacts on plants (91.6%) were categorized as minimal or minor in magnitude with no known adverse long-term impact on non-target plant populations, but a few cacti and thistles are affected at moderate (n = 3), major (n = 7) to massive (n = 1) scale. The largest direct impacts are from two agents (Cactoblastis cactorum on native cacti and Rhinocyllus conicus on native thistles), but these introductions would not be permitted today as more balanced attitudes exist to plant biodiversity, driven by both society and the scientific community. Our analysis shows (as far as is known), weed biological control agents have a biosafety track record of >99% of cases avoiding significant non-target impacts on plant populations. Some impacts could have been overlooked, but this seems unlikely to change the basic distribution of very limited adverse effects. Fewer non-target impacts can be expected in future because of improved science and incorporation of wider values. Failure to use biological control represents a significant opportunity cost from the certainty of ongoing adverse impacts from invasive weeds. It is recommended that a simple five-step scale be used to better communicate the risk of consequences from both action (classical biological control) and no

  19. Assessment of virally vectored autoimmunity as a biocontrol strategy for cane toads.

    Directory of Open Access Journals (Sweden)

    Jackie A Pallister

    Full Text Available BACKGROUND: The cane toad, Bufo (Chaunus marinus, is one of the most notorious vertebrate pests introduced into Australia over the last 200 years and, so far, efforts to identify a naturally occurring B. marinus-specific pathogen for use as a biological control agent have been unsuccessful. We explored an alternative approach that entailed genetically modifying a pathogen with broad host specificity so that it no longer caused disease, but carried a gene to disrupt the cane toad life cycle in a species specific manner. METHODOLOGY/PRINCIPAL FINDINGS: The adult beta globin gene was selected as the model gene for proof of concept of autoimmunity as a biocontrol method for cane toads. A previous report showed injection of bullfrog tadpoles with adult beta globin resulted in an alteration in the form of beta globin expressed in metamorphs as well as reduced survival. In B. marinus we established for the first time that the switch from tadpole to adult globin exists. The effect of injecting B. marinus tadpoles with purified recombinant adult globin protein was then assessed using behavioural (swim speed in tadpoles and jump length in metamorphs, developmental (time to metamorphosis, weight and length at various developmental stages, protein profile of adult globin and genetic (adult globin mRNA levels measures. However, we were unable to detect any differences between treated and control animals. Further, globin delivery using Bohle iridovirus, an Australian ranavirus isolate belonging to the Iridovirus family, did not reduce the survival of metamorphs or alter the form of beta globin expressed in metamorphs. CONCLUSIONS/SIGNIFICANCE: While we were able to show for the first time that the switch from tadpole to adult globin does occur in B. marinus, we were not able to induce autoimmunity and disrupt metamorphosis. The short development time of B. marinus tadpoles may preclude this approach.

  20. Biocontrol and Rapid Detection of Food-Borne Pathogens Using Bacteriophages and Endolysins.

    Science.gov (United States)

    Bai, Jaewoo; Kim, You-Tae; Ryu, Sangryeol; Lee, Ju-Hoon

    2016-01-01

    Bacteriophages have been suggested as natural food preservatives as well as rapid detection materials for food-borne pathogens in various foods. Since Listeria monocytogenes-targeting phage cocktail (ListShield) was approved for applications in foods, numerous phages have been screened and experimentally characterized for phage applications in foods. A single phage and phage cocktail treatments to various foods contaminated with food-borne pathogens including E. coli O157:H7, Salmonella enterica, Campylobacter jejuni, Listeria monocytogenes, Staphylococcus aureus, Cronobacter sakazakii, and Vibrio spp. revealed that they have great potential to control various food-borne pathogens and may be alternative for conventional food preservatives. In addition, phage-derived endolysins with high host specificity and host lysis activities may be preferred to food applications rather than phages. For rapid detection of food-borne pathogens, cell-wall binding domains (CBDs) from endolysins have been suggested due to their high host-specific binding. Fluorescence-tagged CBDs have been successfully evaluated and suggested to be alternative materials of expensive antibodies for various detection applications. Most recently, reporter phage systems have been developed and tested to confirm their usability and accuracy for specific detection. These systems revealed some advantages like rapid detection of only viable pathogenic cells without interference by food components in a very short reaction time, suggesting that these systems may be suitable for monitoring of pathogens in foods. Consequently, phage is the next-generation biocontrol agent as well as rapid detection tool to confirm and even identify the food-borne pathogens present in various foods. PMID:27092128

  1. Comparison of real-time PCR and microscopy to evaluate sclerotial colonisation by a biocontrol fungus.

    Science.gov (United States)

    Kim, Tae Gwan; Knudsen, Guy R

    2011-01-01

    The biocontrol agent Trichoderma harzianum colonises sclerotia of the plant pathogenic fungus Sclerotinia sclerotiorum. Plating of sclerotia typically has been used to determine the incidence of mycoparasitism, but does not quantify the extent to which individual sclerotia are colonised. We developed a specific PCR primer/probe set for the green fluorescent protein (GFP)-transformant T. harzianum ThzID1-M3, which exhibited high precision and reproducibility. Quantitative real-time PCR was evaluated along with epifluorescence microscopy and image analysis to investigate dynamics of colonisation of sclerotia in non-sterile soil. Amounts of ThzID1-M3 DNA and S. sclerotiorum DNA from entire individual sclerotia were quantified using real-time PCR. Epifluorescence micrographs were captured from sclerotial thin-section samples, and GFP fluorescence from these was quantified using computer image analysis in order to estimate colonisation on a per-sclerotium basis. As determined by either method, ThzID1-M3 colonised sclerotia in soil, and both methods quantified colonisation dynamics over time. In a separate experiment, colonisation of sclerotia on agar plates was observed using confocal laser scanning microscopy to view the GFP-fluorescing hyphae of ThzID1-M3. This method, while highly labour-intensive, provided high spatial resolution of colonisation dynamics. Thus, each method has advantages: microscopy combined with image analysis can provide useful information on the spatial and temporal dynamics of colonisation, while real-time PCR can provide a more precise assessment of the extent of sclerotial colonisation over time and can more easily be used to sample entire sclerotia. PMID:21530913

  2. Biocontrol Characteristics of Bacillus Species in Suppressing Stem Rot of Grafted Cactus Caused by Bipolaris cactivora

    Directory of Open Access Journals (Sweden)

    Sooil Bae

    2013-03-01

    Full Text Available One of the most important limiting factors for the production of the grafted cactus in Korea is the qualitative and quantitative yield loss derived from stem rots especially caused by Bipolaris cactivora. This study is aimed to develop microbial control agents useful for the control of the bipolaris stem rot. Two bacteria (GA1-23 and GA4-4 selected out of 943 microbial isolates because of their strong antibiotic activity against B. cactivora were identified as Bacillus subtilis and B. amyloliquefaciens, respectively, by the cultural characteristics, Biolog program and 16S rRNA sequencing analyses. Both bacterial isolates significantly inhibited the conidial germination and mycelial growth of the pathogen with no significant difference between the two, of which the inhibitory efficacies varied depending on the cultural conditions such as temperature, nutritional compositions and concentrations. Light and electron microscopy of the pathogen treated with the bacterial isolates showed the inhibition of spore germination with initial malformation of germ tubes and later formation of circle-like vesicles with no hyphal growth and hyphal disruption sometimes accompanied by hyphal swellings and shrinkages adjacent to the bacteria, suggesting their antibiotic mode of antagonistic activity. Control efficacy of B. subtilis GA1-23 and B. amyloliquefaciens GA4-4 on the cactus stem rot were not as high as but comparable to that of fungicide difenoconazole when they were treated simultaneously at the time of pathogen inoculation. All of these results suggest the two bacterial isolates have a good potential to be developed as biocontrol agents for the bipolaris stem rot of the grafted cactus.

  3. Study on the Biocontrol Activities of Trichoderma species in Greengram with Infected Fungal Pathogens

    International Nuclear Information System (INIS)

    Seven species of Trichoderma were isolated from rhizospheric soil sources and studied by cultural morphology and microscopic examinations. In dual plate assay, antifungal effects of seven Trichoderma strains were screened against three plant pathogenic fungi (Fusarium oxysporum, Rhizoctonia solani and Pythium sp.) on PDA medium and T-5 isolate showed a wide percentage of inhibitory effects on target pathogens with PIRG value. All Trichoderma strains exhibited a clear zone formation on minimal synthetic medium supplemented with 1% colloidal chitin. T-2 and T-5 were the best chitinase producer strains. In vitro screening for protease activity, the highest protease producing activity of Trichoderma isolate (T-2) were observed in pH indicator medium after 7 days incubation. In pot trial experiment, only T-5 strain exhibited more fungal suppression efficiency on green gram plant than commercial fungicide, Trisan and the other strains. So, it can be said that the effective strain was T-5 strain only which have been more antifungal producing power on three fungal pathogens than Trisan and the resting strains.

  4. Development of dry gram-negative bacteria biocontrol products and small pilot tests against dry rot

    Science.gov (United States)

    Pseudomonas fluorescens strains S11:P:12, P22:Y:05, and S22:T:04 suppress four important storage potato maladies; dry rot, late blight, pink rot, and sprouting. Studies were designed to identify methods for producing a dried, efficacious biological control product. The strains were evaluated individ...

  5. Isolation and Bio-control Efficacy of Antagonistic Bacteria against Gray Mold Rot of Pepper after Harvest%辣椒采后灰霉病拮抗细菌的分离及生防效果

    Institute of Scientific and Technical Information of China (English)

    谷会; 朱世江; 詹儒林; 谢江辉

    2011-01-01

    For the purpose of reducing pepper postharvest rot caused by gray mold rot, this study was carried out to isolate and screen antagonistic bacteria from pepper, plants. The result showed that 6 strains of antagonist which could effectively control the postharvest gray mold rot of pepper fruits were isolated, among which the strain PL58 showed the most effective antagonizing activity, and its inhibition circle diameter was more than 39 mm after incubated on the medium plate for 3 d, while the inhibition rate to the pathogeny of pepper gray mold rot was 55.1% after incubated for 7 d, and the strain PL58 which had the characteristic of wide-spectrum antagonism was identified as Bacillus sp. Preliminarily. The suspension(1.0×109fu/mL)of the strain showed strong inhibiting effect to mycelial growth and spore germination of the pathogeny, however, its sterile cellular exudates showed no significant inhibiting effect. Further studies showed that the strain had the best antagonizing effect to the gray mold rot up to 78% on pepper fruits and the best bio-control effect was 37.2% after the fruits stored at 25 °C for 25 d. As a result, the strain PL58 could be applied as a bio-control agent to control pepper postharvest gray mold rot.%为了降低辣椒采后由灰霉病引起的腐烂损失,从辣椒植株内分离筛选拮抗菌,结果获得6株对辣椒灰霉病菌有较强抑制作用的内生细菌,其中PL58菌株的抑菌活性最强,在平板上培养3d后抑菌圈直径大于39 mm,培养7d后对辣椒灰霉病菌的抑制率达55.1%,该菌株初步鉴定为芽孢杆菌,其拮抗作用有一定的广谱性.该菌株的发酵液(1.0×109 cfu/mL)对辣椒灰霉病菌菌丝生长和孢子萌发均有较强的抑制作用,但其无菌滤液没有抑制作用.该菌株在辣椒果实上对灰霉病的拮抗效果最高达78%,辣椒果实在25℃下贮藏25 d后该菌株的生防效果仍有37.2%.因此,PL58菌株可以作为一种生防菌应用于辣椒采后灰霉病的防治.

  6. Teste de especificidade hospedeira de Phaedon confinis (Coleoptera, Chrysomelidae, um potencial agente de biocontrole de Senecio brasiliensis (Asteraceae.

    Directory of Open Access Journals (Sweden)

    Julianne Milléo

    2011-07-01

    Abstract. Senecio brasiliensis (Spreng. Less when ingested by cattle and horses, the plant causes seneciosis, a serious poisoning. Due to the great financial losses to cattle ranchers, controlling the plant using insects has become attractive. Systematic survey efforts have revealed that Phaedon confinis Klug causes serious damage to the plant, and may be a great biocontrol agent. The object was to extend the tests of host specificity to 52 plants using 1st larval instar and adult chrysomelid bettles. The insects were submitted to “no-choice” and “multiple-choice” tests. The following results were obtained: “NO-CHOICE” L1 – 52 plants tested: null 90.39%; negligible damage 5.77%; light 1.92%; and normal in only S. brasiliensis 1.92%, where 31.67% of larvae obtained an adult phase. “NO-CHOICE” ADULTS – 46 plants. Null damage was recorded in 82.60%; 13.04% showed negligible damage; 2.17% light; 2.17% normal in S. brasiliensis. The chysomelids oviposited during observation days only on S. brasiliensis leaves. 615 eggs were oviposited with 73.01% viability. “MULTIPLE CHOICE” LARVAE – nine plants tested. 66.67% null; 11.11% weak; 11.11% negligible damage; and 11.11% normal in S. brasiliensis. The results indicate that the normal diet, oviposition, survival and development of P. confinis is restricted to S. brasiliensis and corroborates its potential as a biocontrol agent.

  7. Intracellular trehalose and sorbitol synergistically promoting cell viability of a biocontrol yeast, Pichia anomala, for aflatoxin reduction.

    Science.gov (United States)

    Hua, Sui Sheng T; Hernlem, Bradley J; Yokoyama, Wallace; Sarreal, Siov Bouy L

    2015-05-01

    Pichia anomala (Wickerhamomyces anomalus) WRL-076 was discovered by a visual screening bioassay for its antagonism against Aspergillus flavus. The yeast was shown to significantly inhibit aflatoxin production and the growth of A. flavus. P. anomala is a potential biocontrol agent for reduction of aflatoxin in the food chain. Maintaining the viability of biocontrol agents in formulated products is a great challenge for commercial applications. Four media, NYG, NYGS, NYGT and NYGST are described which support good growth of yeast cells and were tested as storage formulations. Post growth supplement of 5 % trehalose to NYGST resulted in 83 % viable yeast cells after 12 months in cold storage. Intracellular sorbitol and trehalose concentrations were determined by HPLC analysis at the beginning of the storage and at the end of 12 month. Correlation of cell viability to both trehalose and sorbitol suggested a synergistic effect. Bonferroni (Dunn) t Test, Tukey's Studentized Range (HSD) Test and Duncan's Multiple Range Test, all showed that yeast cell viability in samples with both intracellular trehalose and sorbitol were significantly higher than those with either or none, at a 95 % confidence level. DiBAC4(5) and CFDA-AM were used as the membrane integrity fluorescent stains to create a two-color vital staining scheme with red and green fluorescence, respectively. Yeast cells stored in formulations NYG and NYGS with no detectable trehalose, displayed mostly red fluorescence. Yeast cells in NYGST+5T showed mostly green fluorescence.

  8. Phytic acid enhances biocontrol activity of Rhodotorula mucilaginosa against Penicillium expansum contamination and patulin production in apples

    Directory of Open Access Journals (Sweden)

    Qiya eYang

    2015-11-01

    Full Text Available The effect of Rhodotorula mucilaginosa in combination with phytic acid (PA on blue mold decay and patulin contamination of apples was investigated. Results from this study show that different concentrations of PA were effective in reducing the disease incidence of apples and that PA at concentration of 4 μmol/mL, decreased the incidence of blue mold decay in apples from 86.1% to 62.5%, and showed higher control efficacy compared to untreated, control fruit during storage at 20 °C. However, R. mucilaginosa combined with PA (4 μmol/mL showed better control efficacy of blue mold decay than R. mucilaginosa used as single treatment, the disease incidence was reduced to 62.5% and lesion diameter on apples was reduced to 16.59cm. In in vitro experiments, the addition of PA enhanced the biocontrol effect of R. mucilaginosa against the growth of P. expansum and reduced patulin level when compared with either R. mucilaginosa or PA used separately. R. mucilaginosa together with PA, improved the inhibition of patulin production in wounded apples, decreasing the content of patulin by 89.6% compared to the control, under experimental conditions. Both R. mucilaginosa and R. mucilaginosa in combination with PA degraded patulin in vitro. In conclusion, the appropriate combination of R. mucilaginosa and PA may provide an effective biocontrol method for reducing postharvest decay of apples.

  9. Evidence for biotrophic lifestyle and biocontrol potential of dark septate endophyte Harpophora oryzae to rice blast disease.

    Directory of Open Access Journals (Sweden)

    Zhen-Zhu Su

    Full Text Available The mutualism pattern of the dark septate endophyte (DSE Harpophora oryzae in rice roots and its biocontrol potential in rice blast disease caused by Magnaporthe oryzae were investigated. Fluorescent protein-expressing H. oryzae was used to monitor the colonization pattern. Hyphae invaded from the epidermis to the inner cortex, but not into the root stele. Fungal colonization increased with root tissue maturation, showing no colonization in the meristematic zone, slight colonization in the elongation zone, and heavy colonization in the differentiation zone. H. oryzae adopted a biotrophic lifestyle in roots accompanied by programmed cell death. Real-time PCR facilitated the accurate quantification of fungal growth and the respective plant response. The biocontrol potential of H. oryzae was visualized by inoculation with eGFP-tagged M. oryzae in rice. H. oryzae protected rice from M. oryzae root invasion by the accumulation of H2O2 and elevated antioxidative capacity. H. oryzae also induced systemic resistance against rice blast. This systemic resistance was mediated by the OsWRKY45-dependent salicylic acid (SA signaling pathway, as indicated by the strongly upregulated expression of OsWRKY45. The colonization pattern of H. oryzae was consistent with the typical characteristics of DSEs. H. oryzae enhanced local resistance by reactive oxygen species (ROS and high antioxidative level and induced OsWRKY45-dependent SA-mediated systemic resistance against rice blast.

  10. Potential of chitinolytic Serratia marcescens strain JPP1 for biological control of Aspergillus parasiticus and aflatoxin.

    Science.gov (United States)

    Wang, Kai; Yan, Pei-Sheng; Cao, Li-Xin; Ding, Qing-Long; Shao, Chi; Zhao, Teng-Fei

    2013-01-01

    Serratia marcescens strain JPP1 was isolated from peanut hulls in Huai'an city, Jiangsu Province, China. Its potential to inhibit the mycelial growth of Aspergillus parasiticus and the subsequent aflatoxin production was evaluated. The strain JPP1 could produce chitinase to degrade fungal cell walls, which was the main mechanism of strain JPP1 for biocontrol. Scanning electron microscopy of fungi treated with the crude chitinase revealed abnormal morphological changes. While the strain was grown in the peanut hulls-based medium, the chitinase activity reached 7.39 units. RT-PCR analysis showed that the crude chitinase repressed the transcription of genes involved in the aflatoxin gene cluster, such as aflR, aflC (pksL1), and aflO (dmtA) genes. By visual agar plate assay and tip culture method, the strain JPP1 exhibited remarkable inhibitory effect on mycelia growth (antifungal ratio >95%) and subsequent aflatoxin production (antiaflatoxigenic ratio >98%). An in vitro assay with seed coating agent of bacterial suspension showed that strain JPP1 effectively reduced fungal growth and subsequent aflatoxin production on peanut seeds, and its antagonistic effect was superior to the common agricultural fungicide of carbendazim. These characteristics suggest that S. marcescens JPP1 strain could potentially be utilized for the biological control of phytopathogenic fungi and aflatoxin in Chinese peanut main producing areas.

  11. CHARACTERIZATION OF SOIL TRICHODERMA ISOLATES FOR POTENTIAL BIOCONTROL OF PLANT PATHOGENS

    Directory of Open Access Journals (Sweden)

    S. Matei

    2011-12-01

    Full Text Available Various fungal strains belonging to genus Trichoderma act as biological control agents for soil born plant pathogens. Two new strains of Trichoderma harzianum (T.h. and Trichoderma viride (T.v. were isolated from forest soils in Ilfov county and their morphological aspects, enzymatic and antagonistic activity were examined. Current chemical fungicides had constantly, in time, less influence on pathogens due to their diversity, adaptability and increasing resistance.The paper present the morphological characterization of two strains of Trichoderma isolated from forest soils. Growth rate was higher in strain T.v.SP456 (0,675mm/h than in strain T.h.P8 (0,505mm/h when fungi were grown on Czapek culture medium.Morphological description is completed with photographs of colonies in Petri plates and microscopical aspects of fungal structures belonging to Trichoderma strains SP456 and P8.Comparative aspects concerning the level of main enzymes released by T.h. isolate P8 and T.v.SP456 in liquid culture media showed differences as a function of genetic structure of each fungal isolate. The optimum culture media for inducing peroxidase, polyphenol-oxidase, β-1,3-glucanase activity in T.v.SP456 isolate was Czapek and PDA for phenil-alanin-ammonium-oxidase and chitinase. T.v.SP456 was more efficient than T.h.P8 concerning enzymes activity.The interaction between Trichoderma fungal strains SP456 and P8 and strawberry plant pathogen strains, three belonging to Botrytis cinerea (S1, P1, P2 and one to Phytophtora spp. were examined, also. Both Trichoderma strains act as mycoparasites for plant pathogens. The inhibition percent of radial growth was higher for T.v.SP456 when compared with T.h.P8 for almost all pathogenic isolates.

  12. RNA interference of endochitinases in the sugarcane endophyte Trichoderma virens 223 reduces its fitness as a biocontrol agent of pineapple disease.

    Science.gov (United States)

    Romão-Dumaresq, Aline S; de Araújo, Welington Luiz; Talbot, Nicholas J; Thornton, Christopher R

    2012-01-01

    The sugarcane root endophyte Trichoderma virens 223 holds enormous potential as a sustainable alternative to chemical pesticides in the control of sugarcane diseases. Its efficacy as a biocontrol agent is thought to be associated with its production of chitinase enzymes, including N-acetyl-ß-D-glucosaminidases, chitobiosidases and endochitinases. We used targeted gene deletion and RNA-dependent gene silencing strategies to disrupt N-acetyl-ß-D-glucosaminidase and endochitinase activities of the fungus, and to determine their roles in the biocontrol of soil-borne plant pathogens. The loss of N-acetyl-ß-D-glucosaminidase activities was dispensable for biocontrol of the plurivorous damping-off pathogens Rhizoctonia solani and Sclerotinia sclerotiorum, and of the sugarcane pathogen Ceratocystis paradoxa, the causal agent of pineapple disease. Similarly, suppression of endochitinase activities had no effect on R. solani and S. sclerotiorum disease control, but had a pronounced effect on the ability of T. virens 223 to control pineapple disease. Our work demonstrates a critical requirement for T. virens 223 endochitinase activity in the biocontrol of C. paradoxa sugarcane disease, but not for general antagonism of other soil pathogens. This may reflect its lifestyle as a sugarcane root endophyte. PMID:23110120

  13. Superoxide anion and hydrogen peroxide in the yeast antagonist-fruit interaction: a new role for reactive oxygen species in postharvest biocontrol?

    Science.gov (United States)

    The importance of reactive oxygen species (ROS) in plant defense responses against certain pathogens is well documented. There is some evidence that microbial biocontrol agents also induce a transient production of ROS in a host plant which triggers local and systemic defense responses to pathogens....

  14. Role of 2-hexyl, 5-propyl resorcinol production by Pseudomonas chlororaphis PCL1606 in the multitrophic interactions in the avocado rhizosphere during the biocontrol process.

    Science.gov (United States)

    Calderón, Claudia E; de Vicente, Antonio; Cazorla, Francisco M

    2014-07-01

    Different bacterial traits can contribute to the biocontrol of soilborne phytopathogenic fungus. Among others, (1) antagonism, (2) competition for nutrients and niches, (3) induction of systemic resistance of the plants and (4) predation and parasitism are the most studied. Pseudomonas chlororaphis PCL1606 is an antagonistic rhizobacterium that produces the antifungal metabolite 2-hexyl, 5-propyl resorcinol (HPR). This bacterium can biologically control the avocado white root rot caused by Rosellinia necatrix. Confocal laser scanning microscopy of the avocado rhizosphere revealed that this biocontrol bacterium and the fungal pathogen compete for the same niche and presumably also for root exudate nutrients. The use of derivative mutants in the geners related to HPR biosynthesis (dar genes) revealed that the lack of HPR production by P. chlororaphis PCL1606 negatively influences the bacterial colonisation of the avocado root surface. Microscopical analysis showed that P. chlororaphis PCL1606 closely interacts and colonises the fungal hyphae, which may represent a novel biocontrol mechanism in this pseudomonad. Additionally, the presence of HPR-producing biocontrol bacteria negatively affects the ability of the fungi to infect the avocado root. HPR production negatively affects hyphal growth, leading to alterations in the R. necatrix physiology visible under microscopy, including the curling, vacuolisation and branching of hyphae, which presumably affects the colonisation and infection abilities of the fungus. This study provides the first report of multitrophic interactions in the avocado rhizosphere, advancing our understanding of the role of HPR production in those interactions.

  15. Regulation of aflatoxin biosynthesis and branched-chain amino acids metabolism in Aspergillus flavus by 2-phenylethanol reveal biocontrol mechanism of Pichia anomala

    Science.gov (United States)

    Pichia anomala WRL-076 is a biocontrol yeast which has been shown to inhibit growth and aflatoxin production of A. flavus. Using the SPME-GC/MS analysis we identified that the volatile, 2-phenylethanol (2-PE) produced by this yeast and demonstrated that the compound inhibited aflatoxin production. W...

  16. Polysaccharide Production Benefits Dry Storage Survival of the Biocontrol Agent Pseudomonas fluorescens S11:P:12 Effective Against Several Maladies of Stored Potatoes

    Science.gov (United States)

    Pseudomonas fluorescens S11:P:12 (NRRL B-21133) is a biological control agent able to suppress several potato diseases and sprouting. Notably, it produces a polysaccharide during liquid cultivation; and the objective of this work was to determine the role of this material in the bio-control process...

  17. RNA interference of endochitinases in the sugarcane endophyte Trichoderma virens 223 reduces its fitness as a biocontrol agent of pineapple disease.

    Directory of Open Access Journals (Sweden)

    Aline S Romão-Dumaresq

    Full Text Available The sugarcane root endophyte Trichoderma virens 223 holds enormous potential as a sustainable alternative to chemical pesticides in the control of sugarcane diseases. Its efficacy as a biocontrol agent is thought to be associated with its production of chitinase enzymes, including N-acetyl-ß-D-glucosaminidases, chitobiosidases and endochitinases. We used targeted gene deletion and RNA-dependent gene silencing strategies to disrupt N-acetyl-ß-D-glucosaminidase and endochitinase activities of the fungus, and to determine their roles in the biocontrol of soil-borne plant pathogens. The loss of N-acetyl-ß-D-glucosaminidase activities was dispensable for biocontrol of the plurivorous damping-off pathogens Rhizoctonia solani and Sclerotinia sclerotiorum, and of the sugarcane pathogen Ceratocystis paradoxa, the causal agent of pineapple disease. Similarly, suppression of endochitinase activities had no effect on R. solani and S. sclerotiorum disease control, but had a pronounced effect on the ability of T. virens 223 to control pineapple disease. Our work demonstrates a critical requirement for T. virens 223 endochitinase activity in the biocontrol of C. paradoxa sugarcane disease, but not for general antagonism of other soil pathogens. This may reflect its lifestyle as a sugarcane root endophyte.

  18. Biocontrol traits of plant growth suppressive arbuscular mycorrhizal fungi against root rot in tomato caused by Pythium aphanidermatum

    DEFF Research Database (Denmark)

    Larsen, John; Graham, James H.; Cubero, Jaime;

    2012-01-01

    Arbuscular mycorrhizal (AM) fungi known to cause plant growth depressions in tomato were examined for their biocontrol effects against root rot caused by Pythium aphanidermatum. The main hypothesis was that plant growth suppressive AM fungi would elicit a defence response in the host plant reduci...

  19. What magnitude are observed non-target impacts from weed biocontrol?

    Directory of Open Access Journals (Sweden)

    David Maxwell Suckling

    Full Text Available A systematic review focused by plant on non-target impacts from agents deliberately introduced for the biological control of weeds found significant non-target impacts to be rare. The magnitude of direct impact of 43 biocontrol agents on 140 non-target plants was retrospectively categorized using a risk management framework for ecological impacts of invasive species (minimal, minor, moderate, major, massive. The vast majority of agents introduced for classical biological control of weeds (>99% of 512 agents released have had no known significant adverse effects on non-target plants thus far; major effects suppressing non-target plant populations could be expected to be detectable. Most direct non-target impacts on plants (91.6% were categorized as minimal or minor in magnitude with no known adverse long-term impact on non-target plant populations, but a few cacti and thistles are affected at moderate (n = 3, major (n = 7 to massive (n = 1 scale. The largest direct impacts are from two agents (Cactoblastis cactorum on native cacti and Rhinocyllus conicus on native thistles, but these introductions would not be permitted today as more balanced attitudes exist to plant biodiversity, driven by both society and the scientific community. Our analysis shows (as far as is known, weed biological control agents have a biosafety track record of >99% of cases avoiding significant non-target impacts on plant populations. Some impacts could have been overlooked, but this seems unlikely to change the basic distribution of very limited adverse effects. Fewer non-target impacts can be expected in future because of improved science and incorporation of wider values. Failure to use biological control represents a significant opportunity cost from the certainty of ongoing adverse impacts from invasive weeds. It is recommended that a simple five-step scale be used to better communicate the risk of consequences from both action (classical biological

  20. Greenhouse evaluation of rhizobia as biocontrol agent of root-infecting fungi in okra

    OpenAIRE

    Siddiqui, I. A.; S. Ehteshamul-Haque; Zaki, M. J.; Ghaffar, A.

    2013-01-01

    Nine rhizobial strains isolated from the root nodules of Cicer arietinum, Vigna radiata, V. mungo, Samania saman, Sesbania sesban, Leucinia sp., Prosopis cineraria and Medicago sativa were used to study their effects on root-infecting fungi viz., Macrophomina phaseolina, Fusarium solani and Rhizoctonia solani. In dual culture plate assay, strains of Bradyrhizobium sp., and R. meliloti were found to inhibit radial growth of M. phaseolina, F. solani and R. solani producing zones of inhibition. ...

  1. CHARACTERIZATION OF SOIL TRICHODERMA ISOLATES FOR POTENTIAL BIOCONTROL OF PLANT PATHOGENS

    OpenAIRE

    S. Matei; Gabi-Mirela Matei; Petruta Cornea; Gabriela Popa

    2011-01-01

    Various fungal strains belonging to genus Trichoderma act as biological control agents for soil born plant pathogens. Two new strains of Trichoderma harzianum (T.h.) and Trichoderma viride (T.v.) were isolated from forest soils in Ilfov county and their morphological aspects, enzymatic and antagonistic activity were examined. Current chemical fungicides had constantly, in time, less influence on pathogens due to their diversity, adaptability and increasing resistance.The paper present the mor...

  2. Biocontrol of Penicillium nordicum Growth and Ochratoxin A Production by Native Yeasts of Dry Cured Ham

    OpenAIRE

    Marco Camardo Leggieri; Paola Battilani; Silvia Formenti; Tania Toscani; Nicoletta Simoncini; Roberta Virgili

    2012-01-01

    Twelve yeast strains isolated from the surface of Italian typical dry-cured hams, belonging to D. hansenii, D. maramus, C. famata, C. zeylanoides and H. burtonii species, and previously selected for their ability to grow in dry-cured ham-like substrates, were screened for antagonistic activity against a toxigenic strain of P. nordicum and inhibition of ochratoxin A (OTA) biosynthesis. On average, yeast inhibitory activity was lowered by increasing fungal inoculum and enhanced by NaCl presence...

  3. Seleção de bactérias endofíticas de tomateiro como potenciais agentes de biocontrole e de promoção de crescimento Screening of endophytic bacteria isolated from tomato plants as potencial biocontrol agents and growth promotion

    Directory of Open Access Journals (Sweden)

    Patrícia Baston Barretti

    2009-01-01

    Full Text Available Quarenta isolados bacterianos endofíticos de plantas sadias de tomateiro foram avaliados quanto à sua potencialidade como agentes de biocontrole de doenças do tomateiro. Foi realizada, em casa de vegetação, uma seleção massal utilizando-se Pseudomonas syringae pv. tomato e Alternaria solani, como patógenos desafiantes. Com base na média do número de lesões por planta, quatro isolados foram selecionados como potenciais agentes de biocontrole dessas enfermidades fúngica e bacteriana do tomateiro. Esses isolados foram identificados, por meio do sequenciamento do gene 16S do DNA ribossômico, como Acinetobacter johnsonii (UFV-E05, Serratia marcescens (UFV-E13, Sinorhizobium sp. (UFV-E25 e Bacillus megaterium (UFV-E26. Os mesmos isolados selecionados para o biocontrole também foram avaliados quanto à sua capacidade de promover o crescimento em plantas e somente S. marcescens (UFV-E13 proporcionou aumento na altura das plantas.Forty isolates of endophytic bacteria obtained from healthy tomato plants were tested for their potential as biocontrol agents of tomato diseases. A massal screening was performed at greenhouse using Pseudomonas syringae pv. tomato and Alternaria solani as challenging pathogens. Based on the average number of lesions per plant, four isolates were selected as potential agents of biocontrol of these tomato diseases caused by fungi and bacteria. These isolates were identified by 16S ribosomal DNA sequence analysis as Acinetobacter johnsonii (UFV-E05, Serratia marcescens (UFV-E13, Sinorhizobium sp. (UFV-E25 and Bacillus megaterium (UFV-E26. The four endophytes selected for biocontrol were also evaluated for their ability of promoting plant growth and only S. marcescens (UFV-E13 presented increase in the height of the plants.

  4. Geobacteraceae strains and methods

    Energy Technology Data Exchange (ETDEWEB)

    Lovley, Derek R.; Nevin, Kelly P.; Yi, Hana

    2015-07-07

    Embodiments of the present invention provide a method of producing genetically modified strains of electricigenic microbes that are specifically adapted for the production of electrical current in microbial fuel cells, as well as strains produced by such methods and fuel cells using such strains. In preferred embodiments, the present invention provides genetically modified strains of Geobacter sulfurreducens and methods of using such strains.

  5. Identification of Volatile Metabolites from Fungal Endophytes with Biocontrol Potential towards Fusarium oxysporum F. sp. cubense Race 4

    Directory of Open Access Journals (Sweden)

    A. S. Y. Ting

    2010-01-01

    Full Text Available Problem statement: Fungal endophytes are widely studied for their potential as biocontrol agents towards fungal pathogens. In vitro assessments usually reveal their antibiosis and mycoparasitism nature, but little is understood regarding their production of volatile metabolites as mechanisms of antagonism. Approach: This study explored the potential of fungal endophytes in controlling the pathogen responsible for Fusarium wilt disease. Nine fungal endophytes were tested for their ability to inhibit the growth of the pathogenic Fusarium oxysporum F. sp. cubense race 4 (FocR4 via production of volatile inhibitory metabolites. The type of volatile metabolites produced were subsequently characterized and identified using the Gas-Chromatography Mass-Spectrophotometry (GCMS. Results: Eight of the isolates (BTF05, BTF07, BTF08, BTF15, BTF21, WAA03, WAA02, MIF01 showed positive results with percentages of inhibition varying from 1.43-31.43% while one isolate (ALF01, showed negative result (0% inhibition. Volatile profiles showed that these fungal endophytes produced between 15-47 volatile metabolites per isolate. However, the more volatile metabolites produced by a single endophyte does not indicate better biocontrol potential. Isolate BTF05 produced 47 different volatile metabolites, but has only 8.57% inhibition, compared to isolate BTF21 with 15 metabolites but a percentage of 11.43% inhibition. The potency of the volatile metabolites produced may also influenced the biocontrol potential of the fungal endophytes as some isolates such as BTF08 and MIF01 have only two to three known inhibitory metabolites but have higher PIDG values at 31.43 and 11.43%, respectively. Contrary, isolates WAA02 and WAA03 which has five to six metabolites but PIDG values of less than 3%. Conclusion: Fungal endophytes have the ability to produce several types of volatile metabolites to inhibit the growth of FocR4. These volatile inhibitory metabolites can be further

  6. Combinative effects of a bacterial type-III effector and a biocontrol bacterium on rice growth and disease resistance

    Indian Academy of Sciences (India)

    Haiying Ren; Ganyu Gu; Juying Long; Qian Yin; Tingquan Wu; Tao Song; Shujian Zhang; Zhiyi Chen; Hansong Dong

    2006-12-01

    Expression of HpaGXoo, a bacterial type-III effector, in transgenic plants induces disease resistance. Resistance also can be elicited by biocontrol bacteria. In both cases, plant growth is often promoted. Here we address whether biocontrol bacteria and HpaGXoo can act together to provide better results in crop improvement. We studied effects of Pseudomonas cepacia on the rice variety R109 and the hpaGXoo-expressing rice line HER1. Compared to R109, HER1 showed increased growth, grain yield, and defense responses toward diseases and salinity stress. Colonization of roots by P. cepacia caused 20% and 13% increase, in contrast to controls, in root growth of R109 and HER1. Growth of leaves and stems also increased in R109 but that of HER1 was inhibited. When P. cepacia colonization was subsequent to plant inoculation with Rhizoctonia solani, a pathogen that causes sheath blight, the disease was less severe than controls in both R109 and HER1; HER1, nevertheless, was more resistant, suggesting that P. cepacia and HpaGXoo cooperate in inducing disease resistance. Several genes that critically regulate growth and defense behaved differentially in HER1 and R109 while responding to P. cepacia. In R109 leaves, the OsARF1 gene, which regulates plant growth, was expressed in consistence with growth promotion by P. cepacia. Inversely, OsARF1 expression was coincident with inhibition in growth of HER1 leaves. In both plants, the expression of OsEXP1, which encodes an expansin protein involved in plant growth, was concomitant with growth promotion in leaves instead of roots, in response to P. cepacia. We also studied OsMAPK, a gene that encodes a mitogen-activated protein kinase and controls defense responses toward salinity and infection by pathogens in rice. In response to P. cepacia, an early expression of OsMAPK was coincident with R109 resistance to the disease, while HER1 expressed the gene similarly whether P. cepacia was present or not. Evidently, P. cepacia and GXoo

  7. Characterization of streptomycetes with potential to promote plant growth and biocontrol Caracterização de estreptomicetos com potencial para promoção de crescimento de plantas e biocontrole

    Directory of Open Access Journals (Sweden)

    Carla da Silva Sousa

    2008-02-01

    Full Text Available Studies with streptomycetes in biocontrol programs and plant growth promotion are presented as technological alternatives for environmental sustainable production. This work has the objective of characterizing six isolates of streptomycetes aiming the production of extracellular enzymes, indole acetic acid, capacity for phosphate solubilization, root colonization and growth under different pH and salinity levels. For detection of enzyme activity the isolates were grown in culture media with the enzyme substrates as sole carbon source. The root colonization assay was performed on tomato seedlings grown on 0.6% water-agar medium. Growth under different pH and salinity levels was evaluated in AGS medium with 1%, 1.5%, 2%, 2.5%, and 3% NaCl, and pH levels adjusted to 5.0, 5.5, 6.0, 6.5, and 7.0. All isolates produced the enzymes amylase, catalase, and lipase, as well as indole acetic acid. With one exception (AC-92, all isolates presented cellulolytic and chitinolytic activity, and only AC-26 did not show xylanolytic activity. The isolates AC-147, AC-95, and AC-29 were the highest producers of siderophores. The isolates AC-26 and AC-29 did not show capacity for phosphate solubilization. All isolates colonized tomato roots in vitro, and AC-92 grew under all pH and salinity levels tested. The streptomycetes tested were considered as potential biocontrol and plant growth promotion agents.Estudos com estreptomicetos em programas de biocontrole e promoção de crescimento de plantas são alternativas tecnológicas de produção sustentáveis ao ambiente. O presente trabalho teve como objetivo caracterizar seis isolados de estreptomicetos quanto à produção de enzimas extracelulares, ácido indolacético e sideróforos, capacidade de solubilização de fosfatos, colonização radicular e de crescimento em diferentes níveis de pH e salinidade. Para detectar a produção das enzimas extracelulares, os isolados foram cultivados em meio de cultura contendo

  8. Production of Proteasome Inhibitor Syringolin A by the Endophyte Rhizobium sp. Strain AP16

    Science.gov (United States)

    Bigler, Laurent; Dudler, Robert

    2014-01-01

    Syringolin A, the product of a mixed nonribosomal peptide synthetase/polyketide synthase encoded by the syl gene cluster, is a virulence factor secreted by certain Pseudomonas syringae strains. Together with the glidobactins produced by a number of beta- and gammaproteobacterial human and animal pathogens, it belongs to the syrbactins, a structurally novel class of proteasome inhibitors. In plants, proteasome inhibition by syringolin A-producing P. syringae strains leads to the suppression of host defense pathways requiring proteasome activity, such as the ones mediated by salicylic acid and jasmonic acid. Here we report the discovery of a syl-like gene cluster with some unusual features in the alphaproteobacterial endophyte Rhizobium sp. strain AP16 that encodes a putative syringolin A-like synthetase whose components share 55% to 65% sequence identity (72% to 79% similarity) at the amino acid level. As revealed by average nucleotide identity (ANI) calculations, this strain likely belongs to the same species as biocontrol strain R. rhizogenes K84 (formely known as Agrobacterium radiobacter K84), which, however, carries a nonfunctional deletion remnant of the syl-like gene cluster. Here we present a functional analysis of the syl-like gene cluster of Rhizobium sp. strain AP16 and demonstrate that this endophyte synthesizes syringolin A and some related minor variants, suggesting that proteasome inhibition by syrbactin production can be important not only for pathogens but also for endophytic bacteria in the interaction with their hosts. PMID:24727275

  9. Biocontrol potential of salinity tolerant mutants of Trichoderma harzianum against Fusarium oxysporum Potencial de biocontrole de mutantes sal-tolerantes de Trichoderma harzianum contra Fusarium oxysporum

    Directory of Open Access Journals (Sweden)

    Hassan Abdel-Latif A. Mohamed

    2006-06-01

    Full Text Available Exposing a wild-type culture of Trichoderma harzianum to gamma irradiation induced two stable salt-tolerant mutants (Th50M6 and Th50M11. Under saline conditions, both mutants greatly surpassed their wild type strain in growth rate, sporulation and biological proficiency against Fusarium oxysporum, the causal agent of tomato wilt disease. Tolerant T. harzianum mutants detained a capability to grow and convinced sporulation in growth media containing up to 69 mM NaCl. In comparison with their parent strain, characterization of both mutants confirmed that they have reinforced contents of proline and hydroxyproline, relatively higher sodium content compared to potassium, calcium or magnesium contents, higher level of total phenols. Electrophoretic analysis of total soluble proteins in the salt tolerance mutant Th50M6 showed different bands accumulated in response to 69 mM NaCl. Data also showed that mutants produce certain active metabolites, such as chitinases, cellulases, beta-galactosidases, as well as, some antibiotics i.e., trichodermin, gliotoxin and gliovirin. Trichoderma mutants significantly reduced wilt disease incidence and improved yield and mineral contents of tomato plants under both saline and non-saline soil conditions, as well as, under infested and natural conditions. T. harzianum mutants were also more efficient in dropping the F. oxysporum growth in rhizosphere compared to the wild type strain. Population density of both mutants in rhizosphere far exceeded that of T. harzianum wild type strain.A exposição de uma cepa selvagem de Trichoderma harzianum à irradiação gama induziu dois mutantes tolerantes a sal (Th50M6 e Th50M11. Em condições salinas, os dois mutantes foram muito superiores à cepa selvagem em relação à velocidade de multiplicação, esporulação e eficiência contra Fusarium oxysporum, o agente causador da doença wilt do tomate. Os mutantes tolerantes foram capazes de multiplicação e esporulação em

  10. Biocontrol de la pudrición de raíz de nochebuena de interior con Trichoderma spp. Root rot biocontrol for indoor poinsettia with Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    Felipe de Jesús Osuna-Canizalez

    2012-06-01

    Full Text Available En Morelos, la pudrición de la raíz causada por Fusarium spp., es una de las principales enfermedades de la nochebuena de interior. Por su efecto devastador, en su prevención o control se realizan aplicaciones frecuentes de productos químicos, con los riesgos inherentes a la salud humana y al ambiente. En la búsqueda de alternativas bioracionales al manejo de esta enfermedad, se realizó un ensayo en el que se evaluaron tres cepas comerciales de Trichoderma spp., en tres diferentes sustratos: S1= "tierra de hoja" (70% v/v+tezontle grueso (15% v/v+tezontle fino (10% v/v+agrolita (5% v/v; S2= turba (80% v/v+ fibra de coco (20% v/v; S3= "tierra de hoja" (70% + "tepojal" (30%, en las variedades comerciales más comunes, Freedom Red y Prestige Red. Se utilizó un diseño factorial de tratamientos 4 x 3 x 2 y los tratamientos resultantes se evaluaron en un diseño completamente al azar con seis repeticiones. Respecto a la incidencia de pudrición de la raíz, las cepas comerciales de Trichoderma spp., no mostraron diferencias entre sí ni con el testigo químico. La pudrición de la raíz estuvo asociada con S2, debido a una baja capacidad de aireación, y sólo se presentó en Prestige Red. La población (UFC g-¹ de Trichoderma spp., en el sustrato al término del ciclo, fue igual (pIn Morelos, root rot caused by Fusarium spp., is one of the main diseases of indoor poinsettia. In order to prevent or control its devastating effect, frequent applications of chemical products are performed, with inherent risks to human health and environment. In quest for alternative biorational options, an essay in which three commercial strains of Trichoderma spp., was done, in three different substrates: S1= "organic soil" (70% v/v+thick tezontle (15% v/v+thin tezontle (10% v/v+agrolita (5% v/v; S2= peat moss (80% v/v+ coconut fiber (20% v/v; S3= "organic soil" (70%+"tepojal" (30%, in most common commercial varieties, Freedom Red and Prestige Red. A factorial

  11. Muscle strain (image)

    Science.gov (United States)

    A muscle strain is the stretching or tearing of muscle fibers. A muscle strain can be caused by sports, exercise, a ... something that is too heavy. Symptoms of a muscle strain include pain, tightness, swelling, tenderness, and the ...

  12. Muscle strain treatment

    Science.gov (United States)

    Treatment - muscle strain ... Question: How do you treat a muscle strain ? Answer: Rest the strained muscle and apply ice for the first few days after the injury. Anti-inflammatory medicines or acetaminophen ( ...

  13. Peptaibol, Secondary‐Metabolite, and Hydrophobin Pattern of Commercial Biocontrol Agents Formulated with Species of the Trichoderma harzianum Complex

    DEFF Research Database (Denmark)

    Degenkolb, Thomas; Nielsen, Kristian Fog; Dieckmann, Ralf;

    2015-01-01

    The production of bioactive polypeptides (peptaibiotics) in vivo is a sophisticated adaptation strategy of both mycoparasitic and saprotrophic Trichoderma species for colonizing and defending their natural habitats. This feature is of major practical importance, as the detection of peptaibiotics...... in plant‐protective Trichoderma species, which are successfully used against economically relevant bacterial and fungal plant pathogens, certainly contributes to a better understanding of these complex antagonistic interactions. We analyzed five commercial biocontrol agents (BCAs), namely Canna®, Trichosan......®, Vitalin®, Promot® WP, and TrichoMax®, formulated with recently described species of the Trichoderma harzianum complex, viz. T. afroharzianum, T. simmonsii, and T. guizhouense. By using the well‐established, HPLC/MS‐based peptaibiomics approach, it could unequivocally be demonstrated that all...

  14. Identification of Volatiles Produced by Cladosporium cladosporioides CL-1, a Fungal Biocontrol Agent That Promotes Plant Growth

    Directory of Open Access Journals (Sweden)

    Diby Paul

    2013-10-01

    Full Text Available Certain microbial Volatile Organic Compounds (VOCs have been reported to enhance the growth and development of plants. The biocontrol fungi, Cladosporium cladosporioides CL-1 significantly improved the growth of tobacco seedlings in vitro when they were co-cultivated without physical contact. SPME Quadrupole GC/MS/MS revealed that CL-1 emited the volatiles α-pinene, (−-trans-caryophyllene, tetrahydro-2,2,5,5-tetramethylfuran, dehydroaromadendrene, and (+-sativene. Potential roles of these volatiles in plant growth and development are discussed. Even though there were several fungal VOCs reported in the past that could influence plant growth, their exact mechanisms of action are not fully known. Fungal VOC-mediated plant growth promotion requires in-depth study in order for this technology to be used in large scale for crops, especially those grown under greenhouse conditions.

  15. Identification of volatiles produced by Cladosporium cladosporioides CL-1, a fungal biocontrol agent that promotes plant growth.

    Science.gov (United States)

    Paul, Diby; Park, Kyung Seok

    2013-10-16

    Certain microbial Volatile Organic Compounds (VOCs) have been reported to enhance the growth and development of plants. The biocontrol fungi, Cladosporium cladosporioides CL-1 significantly improved the growth of tobacco seedlings in vitro when they were co-cultivated without physical contact. SPME Quadrupole GC/MS/MS revealed that CL-1 emited the volatiles α-pinene, (-)-trans-caryophyllene, tetrahydro-2,2,5,5-tetramethylfuran, dehydroaromadendrene, and (+)-sativene. Potential roles of these volatiles in plant growth and development are discussed. Even though there were several fungal VOCs reported in the past that could influence plant growth, their exact mechanisms of action are not fully known. Fungal VOC-mediated plant growth promotion requires in-depth study in order for this technology to be used in large scale for crops, especially those grown under greenhouse conditions.

  16. Potencial de la cepa CPA-8 de Bacillus subtilis como agente de biocontrol de enfermedades de postcosecha de fruta

    OpenAIRE

    Yánez Mandizábal, Viviana del Rocío

    2012-01-01

    La limitació en l’ús de fungicides per al control de malalties en postcollita de fruita és una problemàtica d’elevada magnitud en el sector fructícola actual. Degut a això l’ús d’estratègies alternatives com el control biològic microbià són fonamentals per a la producció de fruita de qualitat. Malgrat tot, el desenvolupament de programes de biocontrol eficaços requereix d’un coneixement profund de la capacitat de control i els mecanismes d’acció utilitzats per l’agent microbià ...

  17. Yeasts from native Brazilian Cerrado plants: Occurrence, diversity and use in the biocontrol of citrus green mould.

    Science.gov (United States)

    Sperandio, Eugenio Miranda; do Vale, Helson Mario Martins; Moreira, Geisianny Augusta Monteiro

    2015-11-01

    Yeasts are some of the most important postharvest biocontrol agents. Postharvest oranges frequently deteriorate due to green mould (Penicillium digitatum), which causes significant losses. The aims of this study were to determine the composition and diversity of yeasts on plants of the Brazilian Cerrado and to explore their potential for inhibiting citrus green mould. Leaves and fruit of Byrsonima crassifolia and Eugenia dysenterica were collected from Cerrado conservation areas, and thirty-five yeasts were isolated and identified by sequencing the D1-D2 domain of the rDNA large subunit (26S). The isolates represented the Aureobasidium, Meyerozyma, Candida, and Pichia genera. Three isolates identified as Aureobasidium pullulans exhibited potential for the control of P. digitatum in both in vitro and in vivo tests; these isolates reduced the incidence of disease and increased the storage time of fruit. Aureobasidium. pullulans has immense potential for the biological control of filamentous fungi. PMID:26466874

  18. In-vitro evaluation of fungicides, plant extracts and bio-control agents against rice blast pathogen magnaporthe oryzae couch

    International Nuclear Information System (INIS)

    Among 5 fungicides viz., Thiophanate-methyl, Carbendazim, Fosetyl-aluminium, Mancozeb and Copper oxychloride, used against the Magnaporthe oryzae, only Mancozeb appeared as the highly effective fungicide that completely inhibited the mycelial growth of the fungus. All other fungicides showed little effect at higher concentrations. The extracts of garlic (Allium sativum L.), neem (Azadirachta indica L.) and calatropis (Calotropis procera L.) when used against M. oryzae by food poisoning method, only higher dose of garlic completely inhibited the mycelial growth of the test fungus. Six bio-control agents viz., Trichoderma harzianum, Trichoderma polysporum, Trichoderma pseudokoningii, Gliocladium virens, Paecilomyces variotii and Paecilomyces lilacinus were used. Maximum mycelial inhibition of M. oryzae was provided by P. lilacinus followed by Trichoderma spp. (author)

  19. Different Effects of Metarhizium anisopliae Strains IMI330189 and IBC200614 on Enzymes Activities and Hemocytes of Locusta migratoria L.

    Science.gov (United States)

    Cao, Guangchun; Jia, Miao; Zhao, Xia; Wang, Lei; Tu, Xiongbing; Wang, Guangjun; Nong, Xiangqun; Zhang, Zehua

    2016-01-01

    Background Metarhizium is an important class of entomopathogenic fungi in the biocontrol of insects, but its virulence is affected by insect immunity. To clarify the mechanism in virulence of Metarhizium, we compared the immunological differences in Locusta migratoria L. when exposed to two strains of Metarhizium anisopliae (Ma). Results The virulence of Ma IMI330189 was significantly higher than that of Ma IBC200614 to locust, and IMI330189 overcame the hemocytes and began destroying the hemocytes of locust at 72 h after spray, while locust is immune to IBC200614. IMI330189 could overcome the humoral immunity of locust by inhibiting the activities of phenol oxidase (PO), esterases, multi-function oxidases (MFOs) and acetylcholinesterases in locust while increasing the activities of glutathione-S-transferases (GSTs), catalase and aryl-acylamidase (AA). However IBC200614 inhibit the activities of GSTs and AA in locust and increase the activities of MFOs, PO, superoxide dismutase, peroxidase and chitinase in locust. The changes of enzymes activities in period of infection showed that the time period between the 2nd and the 5th day after spray is critical in the pathogenic process. Conclusion These results found the phenomenon that Ma initiatively broke host hemocytes, revealed the correlation between the virulence of Ma and the changes of enzymes activities in host induced by Ma, and clarified the critical period in the infection of Ma. So, these results should provide guidance for the construction of efficient biocontrol Ma strains. PMID:27227835

  20. Selection, Identification and Optimization of the Growth Water Probiotic Consortium of Mangrove Ecosystems as Bioremediation and Biocontrol in Shrimp Ponds

    Directory of Open Access Journals (Sweden)

    Wilis Ari Setyati

    2015-01-01

    Full Text Available Shrimp aquaculture is an activity that potentially generates organic waste. The accumulation of organic matter is becoming one of the main factors causing the emergence of disease. Problem-solving approach that is most effective is through bioremediation. The aims of this study were to select, identify and cultivate bacteria from mangrove sediments from Cilacap, Rembang and Banyuwangi which potentially as probiotic consortium of bioremediation activity and biocontrol. The results showed that total of 45 isolates (proteolytic, 35 isolates (amylolytic, 35 isolates (lipolytic, and 18 isolates (cellulolytic. There were 59 bacterial isolates had antibacterial activity of vibrio (V. harveyi, V. alginolyticus, V. vulnificus and V. anguilarum. Based on the identification of 16 S-rRNA genes, 4 isolates showed that the C2 isolate was identified as Bacillus subtilis, C11 isolate was identified as Bacillus firmus, C13 and C14 isolates were identified as B. Flexus. This study concluded that cultivation of Bacillus subtilis C2 optimum at 2% molase and yeast extract 0.5% at pH 8 and 30 0C. Bacillus firmus C11 optimum at 2% molase and yeast extract 0.5% at pH 8 and 30 0C. Bacillus flexus C13 optimum at 2% glucose and yeast extract 0.5% at pH 8 and 30 0C. Bacillus flexus C14 optimum at 4% molase and yeast extract 0.25% at pH 8 and 30 0C. The result of culture applications of 4 isolates showed an effect of increasing shrimp weight by 141, 9% compared by the control.Keywords: sediment, mangrove, bioremediation, biocontrol

  1. Chitinase production by Bacillus subtilis ATCC 11774 and its effect on biocontrol of Rhizoctonia diseases of potato.

    Science.gov (United States)

    Saber, Wesam I A; Ghoneem, Khalid M; Al-Askar, Abdulaziz A; Rashad, Younes M; Ali, Abeer A; Rashad, Ehsan M

    2015-12-01

    Stem canker and black scurf of potato, caused by Rhizoctonia solani, can be serious diseases causing an economically significant damage. Biocontrol activity of Bacillus subtilis ATCC 11774 against the Rhizoctonia diseases of potato was investigated in this study. Chitinase enzyme was optimally produced by B. subtilis under batch fermentation conditions similar to those of the potato-growing soil. The maximum chitinase was obtained at initial pH 8 and 30 °C. In vitro, the lytic action of the B. subtilis chitinase was detected releasing 355 μg GlcNAc ml⁻¹ from the cell wall extract of R. solani and suggesting the presence of various chitinase enzymes in the bacterial filtrate. In dual culture test, the antagonistic behavior of B. subtilis resulted in the inhibition of the radial growth of R. solani by 48.1% after 4 days. Moreover, the extracted B. subtilis chitinase reduced the growth of R. solani by 42.3% when incorporated with the PDA plates. Under greenhouse conditions, application of a bacterial suspension of B. subtilis at 109 cell mL⁻¹ significantly reduced the disease incidence of stem canker and black scurf to 22.3 and 30%, respectively. In addition, it significantly improved some biochemical parameters, growth and tubers yield. Our findings indicate two points; firstly, B. subtilis possesses a good biocontrol activity against Rhizoctonia diseases of potato, secondly, the harmonization and suitability of the soil conditions to the growth and activity of B. subtilis guaranteed a high controlling capacity against the target pathogen. PMID:26616375

  2. Biocontrol of Aspergillus species on peanut kernels by antifungal diketopiperazine producing Bacillus cereus associated with entomopathogenic nematode.

    Science.gov (United States)

    Kumar, Sasidharan Nishanth; Sreekala, Sreerag Ravikumar; Chandrasekaran, Dileep; Nambisan, Bala; Anto, Ruby John

    2014-01-01

    The rhabditid entomopathogenic nematode associated Bacillus cereus and the antifungal compounds produced by this bacterium were evaluated for their activity in reducing postharvest decay of peanut kernels caused by Aspergillus species in in vitro and in vivo tests. The results showed that B. cereus had a significant effect on biocontrol effectiveness in in vitro and in vivo conditions. The antifungal compounds produced by the B. cereus were purified using silica gel column chromatography and their structure was elucidated using extensive spectral analyses. The compounds were identified as diketopiperazines (DKPs) [cyclo-(L-Pro-Gly), cyclo(L-Tyr-L-Tyr), cyclo-(L-Phe-Gly) and cyclo(4-hydroxy-L-Pro-L-Trp)]. The antifungal activities of diketopiperazines were studied against five Aspergillus species and best MIC of 2 µg/ml was recorded against A. flavus by cyclo(4-hydroxy-L-Pro-L-Trp). To investigate the potential application of cyclo(4-hydroxy-L-Pro-L-Trp) to eliminate fungal spoilage in food and feed, peanut kernels was used as a food model system. White mycelia and dark/pale green spores of Aspergillus species were observed in the control peanut kernels after 2 days incubation. However the fungal growth was not observed in peanut kernels treated with cyclo(4-hydroxy-L-Pro-L-Trp). The cyclo(4-hydroxy-L-Pro-L-Trp) was nontoxic to two normal cell lines [fore skin (FS) normal fibroblast and African green monkey kidney (VERO)] up to 200 µg/ml in MTT assay. Thus the cyclo(4-hydroxy-L-Pro-L-Trp) identified in this study may be a promising alternative to chemical preservatives as a potential biopreservative agent which prevent fungal growth in food and feed. To the best of our knowledge, this is the first report demonstrating that the entomopathogenic nematode associated B. cereus and cyclo(4-hydroxy-L-Pro-L-Trp) could be used as a biocontrol agents against postharvest fungal disease caused by Aspergillus species.

  3. Biological control as a strategy to reduce the impact of mycotoxins in peanuts, grapes and cereals in Argentina.

    Science.gov (United States)

    Chulze, S N; Palazzini, J M; Torres, A M; Barros, G; Ponsone, M L; Geisen, R; Schmidt-Heydt, M; Köhl, J

    2015-01-01

    Mycotoxins including aflatoxins, deoxynivalenol, fumonisins and ochratoxin A are among the main fungal secondary metabolites detected as natural contaminants in South America in different commodities such as peanuts (aflatoxins), cereals (deoxynivalenol and fumonisins) or grapes (ochratoxin A). Different strategies including crop rotation, tillage practices, fungicide application and planting less susceptible cultivars are used in order to reduce the impact of these mycotoxins in both food and feed chains. The development of fungicide resistance in many fungal pathogens as well as rising of public concern on the risks associated with pesticide use led to the search for alternative environmentally friendly methods. Biological control of plant pathogens and toxigenic fungi offers an alternative that can complement chemical control in the frame of an integrated pest management to reduce the impact of mycotoxins in the food and feed chains. The advances made in Argentina on reducing the impact of toxigenic fungi and mycotoxins in peanut, grapes and cereals using the biocontrol strategy are summarised. Native bacteria, yeasts and filamentous fungi have been selected to evaluate them as potential biocontrol agents. Field trials showed that Bacillus subtilis RC 218 and Brevibacillus sp. RC 263 were effective at reducing deoxynivalenol accumulation in wheat. The application of Clonostachys rosea isolates on wheat stubble reduced Fusarium colonisation on the stubble. Bacillus amyloliquefaciens and Microbacterium oleovorans showed good activity to control both Fusarium verticillioides growth and the accumulation of fumonisins at pre-harvest stage in maize. Control of toxigenic Aspergillus flavus and aflatoxin accumulation in peanuts was achieved using a native atoxigenic Aspergillus flavus strain based on competitive exclusion of the toxigenic strains. Kluyveromyces thermotolerans strains were used as biocontrol agents to reduce the impact of Aspergillus section Nigri and

  4. Genome-scale investigation of phenotypically distinct but nearly clonal Trichoderma strains.

    Science.gov (United States)

    Lange, Claudia; Weld, Richard J; Cox, Murray P; Bradshaw, Rosie E; McLean, Kirstin L; Stewart, Alison; Steyaert, Johanna M

    2016-01-01

    Biological control agents (BCA) are beneficial organisms that are applied to protect plants from pests. Many fungi of the genus Trichoderma are successful BCAs but the underlying mechanisms are not yet fully understood. Trichoderma cf. atroviride strain LU132 is a remarkably effective BCA compared to T. cf. atroviride strain LU140 but these strains were found to be highly similar at the DNA sequence level. This unusual combination of phenotypic variability and high DNA sequence similarity between separately isolated strains prompted us to undertake a genome comparison study in order to identify DNA polymorphisms. We further investigated if the polymorphisms had functional effects on the phenotypes. The two strains were clearly identified as individuals, exhibiting different growth rates, conidiation and metabolism. Superior pathogen control demonstrated by LU132 depended on its faster growth, which is a prerequisite for successful distribution and competition. Genome sequencing identified only one non-synonymous single nucleotide polymorphism (SNP) between the strains. Based on this SNP, we successfully designed and validated an RFLP protocol that can be used to differentiate LU132 from LU140 and other Trichoderma strains. This SNP changed the amino acid sequence of SERF, encoded by the previously undescribed single copy gene "small EDRK-rich factor" (serf). A deletion of serf in the two strains did not lead to identical phenotypes, suggesting that, in addition to the single functional SNP between the nearly clonal Trichoderma cf. atroviride strains, other non-genomic factors contribute to their phenotypic variation. This finding is significant as it shows that genomics is an extremely useful but not exhaustive tool for the study of biocontrol complexity and for strain typing. PMID:27190719

  5. A BROAD-SPECTRUM ANTAGONISTIC ACTIVITY OF THE BIOCONTROL AGENT PSEUDOMONAS SYNXANTHA BG33R

    Science.gov (United States)

    Pseudomonas synxantha strain BG33R was previously reported to be effective in controlling the Peach Tree Short Life (PTSL) syndrome by suppressing the ring nematode (Mesocriconema xenoplax) through terminating the eggs of the second juvenile stage from further development. In this research the spec...

  6. Biocontrol of Fusarium graminearum Growth and Deoxynivalenol Production in Wheat Kernels with Bacterial Antagonists

    Directory of Open Access Journals (Sweden)

    Cuijuan Shi

    2014-01-01

    Full Text Available Fusarium graminearum is the main causal pathogen affecting small-grain cereals, and it produces deoxynivalenol, a kind of mycotoxin, which displays a wide range of toxic effects in human and animals. Bacterial strains isolated from peanut shells were investigated for their activities against F. graminearum by dual-culture plate and tip-culture assays. Among them, twenty strains exhibited potent inhibition to the growth of F. graminearum, and the inhibition rates ranged from 41.41% to 54.55% in dual-culture plate assay and 92.70% to 100% in tip-culture assay. Furthermore, eighteen strains reduced the production of deoxynivalenol by 16.69% to 90.30% in the wheat kernels assay. Finally, the strains with the strongest inhibitory activity were identified by morphological, physiological, biochemical methods and also 16S rDNA and gyrA gene analysis as Bacillus amyloliquefaciens. The current study highlights the potential application of antagonistic microorganisms and their metabolites in the prevention of fungal growth and mycotoxin production in wheat kernels. As a biological strategy, it might avoid safety problems and nutrition loss which always caused by physical and chemical strategies.

  7. Controlled release of Pantoea agglomerans E325 for biocontrol of fire blight

    Science.gov (United States)

    Microencapsulation and controlled release of Pantoea agglomerans strain E325 (E325), which is an antagonist to bacterial pathogen (Erwinia amylovora) of fire blight, a devastating disease of apple and pear, have been investigated. Uniform core-shell alginate microcapsules (AMCs), 60-300 µm in diamet...

  8. Production of lipopeptides among Bacillus strains showing growth inhibition of phytopathogenic fungi.

    Science.gov (United States)

    Velho, R V; Medina, L F C; Segalin, J; Brandelli, A

    2011-07-01

    The biological activity and the presence of genes sfp and ituD (surfactin and iturin A) among Bacillus strains isolated from the Amazon basin were determined. Bacillus spp. were tested for hemolytic activity and inhibition of fungal growth by agar plate assays in parallel with PCR for identification of sfp and ituD genes. All strains tested produced surface-active compounds, giving evidence by lysis of erythrocytes and emulsifying activity on mineral oil and soybean oil. These strains of Bacillus caused growth inhibition of several phytopathogenic fungi, including Fusarium spp., Aspergillus spp., and Bipolaris sorokiniana. The presence of genes ituD and sfp was confirmed by PCR and sequence analysis. The only exception was Bacillus sp. P34 that lacks sfp gene. Lipopeptides were isolated from culture supernatants and analyzed by mass spectrometry. Characteristic m/z peaks for surfactin and iturin were observed, and some strains also produced fengycin and bacillomycin. The remarkable antifungal activity showed by the strains could be associated with the co-production of three or more lipopeptide antibiotics. Screening for novel bacteria producing useful biosurfactants or biocontrol agents for agriculture is a topic of greatest importance to eliminate chemical pollutants.

  9. Short communication. A review on the efficacy tests and risk analyses conducted on Chondrostereum purpureum, a potential biocontrol agent, in Finland

    Energy Technology Data Exchange (ETDEWEB)

    Hantula, J.; Hamberg, L.; Vartiamaki, H.; Korhonen, K.; Uotila, A.

    2012-11-01

    Hardwood sprouting is a problem in forest regeneration areas, under electric lines, on roadsides and railways. In Finland, isolates of Chondrostereum purpureum were screened by field experiments for their efficiency to control sprouting. The proportion of dead stumps with the best isolates exceeded 80% on birch (Betula pendula and B. pubescens), and C. purpureum was also found to affect the sprouting of aspen (Populus tremula) and rowan (Sorbus aucuparia). The risks of C. purpureum based biocontrol were evaluated by population genetic analysis. It showed that C. purpureum is a geographically undifferentiated species that does not reproduce clonally. The risk of infection of non-target trees was found to be highest in early spring. These findings suggest that the risks of using C. purpureum in biocontrol are small. (Author) 36 refs.

  10. Program Calibrates Strain Gauges

    Science.gov (United States)

    Okazaki, Gary D.

    1991-01-01

    Program dramatically reduces personnel and time requirements for acceptance tests of hardware. Data-acquisition system reads output from Wheatstone full-bridge strain-gauge circuit and calculates strain by use of shunt calibration technique. Program nearly instantaneously tabulates and plots strain data against load-cell outputs. Modified to acquire strain data for other specimens wherever full-bridge strain-gauge circuits used. Written in HP BASIC.

  11. 西藏发酵制品中拮抗酵母对水晶梨青霉病的防治效果研究%Evaluation of yeasts from Tibetan fermented products as agents for biocontrol of blue mold of Nashi pear fruits

    Institute of Scientific and Technical Information of China (English)

    Hao HU; Yang XU; Huang-ping LU; Rui XIAO; Xiao-dong ZHENG‡; Ting YU

    2015-01-01

      结论:从西藏发酵制品中筛选得到一株拮抗酵母 R. mucilaginosa,其对水晶梨采后青霉病具有较好的生物防治效果。研究结果发现其主要作用机理为在果实伤口处与病原菌的营养与空间竞争。通过进一步研究,可开发为梨果实采后病害防治的新型生物保鲜剂。%A total of 20 strains of yeast isolated from Tibetan fermented products were screened for antagonism against blue mold of pear caused by Penicil ium expansum. Six isolates that inhibited incidence of postharvest decay by 35%or more were selected for further screening. Among them, the most effective was Rhodotorula mucilaginosa. The results showed that washed cel suspensions of R. mucilaginosa yielded better antagonistic efficacy than un-washed cel-culture mixtures, cel-free culture filtrates, and autoclaved cell cultures. Biocontrol activity improved with increasing concentrations of incubated cel s. The best concentration was 1×108 cel s/ml, at which the incidence of decay was only 16.7% after 6 d of incubation. The germination of conidia of P. expansum in vitro was significantly inhibited by both washed cell-suspensions and unwashed cel-culture mixtures. Rapid colonization by yeast at different concentrations showed a relationship between yeast-cel concentration and biocontrol activity. Although the titratable acidity of pear fruits increased after treatment, R. mucilaginosa did not affect the total soluble solids or ascorbic acid content. This is the first study to report that the yeast R. mucilaginosa from Tibet Autonomous Region of China may have potential as an antagonist to control the postharvest decay of pear fruits.

  12. RNA Interference of Endochitinases in the Sugarcane Endophyte Trichoderma virens 223 Reduces Its Fitness as a Biocontrol Agent of Pineapple Disease

    OpenAIRE

    Aline S Romão-Dumaresq; Welington Luiz Araújo; Nicholas J Talbot; Thornton, Christopher R.

    2012-01-01

    The sugarcane root endophyte Trichoderma virens 223 holds enormous potential as a sustainable alternative to chemical pesticides in the control of sugarcane diseases. Its efficacy as a biocontrol agent is thought to be associated with its production of chitinase enzymes, including N-acetyl-ß-D-glucosaminidases, chitobiosidases and endochitinases. We used targeted gene deletion and RNA-dependent gene silencing strategies to disrupt N-acetyl-ß-D-glucosaminidase and endochitinase activities of t...

  13. In Vitro Evaluation Of Selected Plant Extracts As Biocontrol Agents Against Black Mold Aspergillus Niger Van Tieghem Of Onion Bulbs Allium Cepa L.

    OpenAIRE

    SAIFELDIN A. F. EL-NAGERABI; Awad H. M. Ahmed; ABDULKADIR E. ELSHAFIE

    2015-01-01

    Black mold disease caused by Aspergillus niger V. Tiegh. is the most devastating infection occurs in onions Allium cepa L. under field and store conditions. The use of biocontrol agents is ecofriendly approach for controlling seedborne and soilborne diseases compared to the use of toxic synthetic fungicides. This study has been designed to assess the contamination levels of onion seeds with A. niger and its effect on seed germination and to evaluate the in vitro antifungal activity of Prunus ...

  14. Zoospore Homing and Infection Events: Effects of the Biocontrol Bacterium Burkholderia cepacia AMMDR1 on Two Oomycete Pathogens of Pea (Pisum sativum L.)

    OpenAIRE

    Heungens, K; Parke, J. L.

    2000-01-01

    Burkholderia cepacia AMMDR1 is a biocontrol agent that protects pea and sweet corn seeds from Pythium damping-off in field experiments. The goal of this work was to understand the effect of B. cepacia AMMDR1 on Pythium aphanidermatum and Aphanomyces euteiches zoospore homing events and on infection of pea seeds or roots. In vitro, B. cepacia AMMDR1 caused zoospore lysis, prevented cyst germination, and inhibited germ tube growth of both oomycetes. B. cepacia AMMDR1 also reduced the attractive...

  15. Biocontrol Activity of Bacillus subtilis Isolated from Agaricus bisporus Mushroom Compost Against Pathogenic Fungi.

    Science.gov (United States)

    Liu, Can; Sheng, Jiping; Chen, Lin; Zheng, Yanyan; Lee, David Yue Wei; Yang, Yang; Xu, Mingshuang; Shen, Lin

    2015-07-01

    Bacillus subtilis strain B154, isolated from Agaricus bisporus mushroom compost infected by red bread mold, exhibited antagonistic activities against Neurospora sitophila. Antifungal activity against phytopathogenic fungi was also observed. The maximum antifungal activity was reached during the stationary phase. This antifungal activity was stable over a wide pH and temperature range and was not affected by proteases. Assay of antifungal activity in vitro indicated that a purified antifungal substance could strongly inhibit mycelia growth and spore germination of N. sitophila. In addition, treatment with strain B154 in A. bisporus mushroom compost infected with N. sitophila significantly increased the yield of bisporus mushrooms. Ultraviolet scan spectroscopy, tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis, matrix-associated laser desorption ionization time-of-flight mass spectrometry, and electrospray ionization tandem mass spectrometry analyses revealed a molecular weight consistent with 1498.7633 Da. The antifungal compound might belong to a new type of lipopeptide fengycin. PMID:26050784

  16. Greenhouse evaluation of rhizobia as biocontrol agent of root-infecting fungi in okra

    Directory of Open Access Journals (Sweden)

    I. A. Siddiqui

    2013-12-01

    Full Text Available Nine rhizobial strains isolated from the root nodules of Cicer arietinum, Vigna radiata, V. mungo, Samania saman, Sesbania sesban, Leucinia sp., Prosopis cineraria and Medicago sativa were used to study their effects on root-infecting fungi viz., Macrophomina phaseolina, Fusarium solani and Rhizoctonia solani. In dual culture plate assay, strains of Bradyrhizobium sp., and R. meliloti were found to inhibit radial growth of M. phaseolina, F. solani and R. solani producing zones of inhibition. Bradyrhizobium sp., and R. meliloti either used as seed dressing or as soil drench significantly suppressed root-rot infection caused by M. phaseolina, F. solani and R. solani in okra, a non-1eguminous crop under greenhouse conditions. Biomass of plants was also higher in the presence of rhizobia.

  17. Biocontrol of Soil Fungi in Tomato with Microencapsulates Containing Bacillus subtilis

    OpenAIRE

    Marcela H. Suarez; Francisco D. Hernandez-Castillo; Gabriel Gallegos-Morales; R. H. Lira-Saldivar; Raul Rodriguez-Herrera; Aguilar, Cristobal N.

    2011-01-01

    Problem statement: An option to reduce pollution by synthetic agro-chemical in root plant disease management is the use of antagonist rhizobacteria belonging to Bacillus genus, because their inhibitory properties, stimulation of plant growth and crop yield increase. Approach: This study was carried out in order to evaluate if Bacillus subtilis strains could play an antagonists role of plant pathogens and if they can be microencapsulated inside a biopolymer matrix. It was adapted an equipment ...

  18. Biocontrol of Rhizoctonia solani, the causal agent of bean damping-off by fluorescent pseudomonads.

    Science.gov (United States)

    Afsharmanesh, H; Ahmadzadeh, M; Sharifi-Tehrani, A

    2006-01-01

    Rhizosphere bacteria belonging to the fluorescent pseudomonads are receiving increasing attention for the protection of plants against soil-borne fungal pathogens. Among these pathogens, Rhizoctonia solani, the causal agent of bean damping- off is very important in bean fields of Iran. In this study, the antagonistic activity of 46 isolates of fluorescent pseudomonads (isolated from different area of Iran) and Pseudomonas fluorescens strain CHA0 investigated against one isolate of R. solani. About 64% of isolates revealed antagonistic activity against R. solani. Production of antifungal metabolites such as HCN, siderophore and protease was evaluated. The results showed that 97.8%, 17% and 78% of isolates produced siderophore, HCN and protease respectively. There was no significant correlation between antagonistic activity and production of these metabolites. Isolates P-5, P-10 and P-32 with strain CHA0 were selected in order to investigate involvement of siderophore, volatile metabolites (HCN), and non-volatile metabolites in reducing mycelial growth of R. olani. Isolate P-5 showed much more inhibitory effect by production of volatile metabolites and siderophore. Non-volatile metabolites in isolates P-32 and P-5 completely inhibited mycelial growth of the fungus. After the primary labrotory tests, isolates P-14, P-35, P-30, P-5 and strain CHA0 were selected for in vivo experiments. These selected isolates with benomyl fungicide were used as seed coating and soil drenching in sterile soil under greenhouse condition. The result indicated that in seed treatment method, isolates P-30 by 66% had the most effect in disease reduction while in soil treatment method, strain CHAO by 60% had the most effect, such that this two isolates showed significant differences in comparison with plants inoculated with R. solani inoculums. PMID:17390854

  19. Biocontrol and plant growth-promoting activity of rhizobacteria from Chinese fields with contaminated soils

    OpenAIRE

    WANG, XUEFEI; Mavrodi, Dmitri V.; Ke, Linfeng; Mavrodi, Olga V; Yang, Mingming; Thomashow, Linda S.; Zheng, Na; Weller, David M; Zhang, Jibin

    2014-01-01

    The aim of this study was to inventory the types of plant growth-promoting rhizobacteria (PGPR) present in the rhizosphere of plants grown in soils contaminated with heavy metals, recalcitrant organics, petroleum sewage or salinity in China. We screened 1223 isolates for antifungal activity and about 24% inhibited R hizoctonia solani or S clerotinia sclerotiorum. Twenty-four strains inhibitory to R . solani, G aeumannomyces graminis var. tritici and/or S . sclerotiorum and representing the do...

  20. Evaluation of antifungal activity of carbonate and bicarbonate salts alone or in combination with biocontrol agents in control of citrus green mold.

    Science.gov (United States)

    Zamani, M; Sharifi Tehrani, A; Ali Abadi, A Alizadeh

    2007-01-01

    The aim of this research was to determine if the attacks of green mold on orange could be reduced by edible salts alone or in combination with biocontrol agent. For this purpose toxicity to Pantoea digitatum and practical use of sodium carbonate (SC), sodium bicarbonate (SBC) and potassium carbonate, and potassium bicarbonate alone or in combination with antagonistic bacteria (Pseudomonas fluorescens isolate PN, Bacillus subtilis isolate VHN, Pantoea agglomerans isolate CA) to control green mold were determined. All were fungistatic. SC and SBC were equal and superior to the other salts for control of green mold on oranges inoculated 6h before treatment and were chosen for subsequent trails under cold storage conditions. The biocontrol agents were found completely tolerant to 3% sodium bicarbonate and sodium carbonate at room temperature; although their culturability was reduced by > 1000-fold after 60 min in 1% other salt solutions. Satisfactory results were also obtained with the combined treatment for control of green mold. A significant increase in biocontrol activity of all isolate was observed when combined with sodium carbonate and sodium bicarbonate. The treatments comprising CA combined with SB was as effective as fungicide treatment. Thus, use of sodium bicarbonate treatment at 3% followed by the antagonist P. agglomerans CA could be an alternative to chemical fungicides for control of green mold on oranges. PMID:18396809

  1. In Planta Biocontrol of Pectobacterium atrosepticum by Rhodococcus erythropolis Involves Silencing of Pathogen Communication by the Rhodococcal Gamma-Lactone Catabolic Pathway.

    Directory of Open Access Journals (Sweden)

    Corinne Barbey

    Full Text Available The virulence of numerous Gram-negative bacteria is under the control of a quorum sensing process based on synthesis and perception of N-acyl homoserine lactones. Rhodococcus erythropolis, a Gram-positive bacterium, has recently been proposed as a biocontrol agent for plant protection against soft-rot bacteria, including Pectobacterium. Here, we show that the γ-lactone catabolic pathway of R. erythropolis disrupts Pectobacterium communication and prevents plant soft-rot. We report the first characterization and demonstration of N-acyl homoserine lactone quenching in planta. In particular, we describe the transcription of the R. erythropolis lactonase gene, encoding the key enzyme of this pathway, and the subsequent lactone breakdown. The role of this catabolic pathway in biocontrol activity was confirmed by deletion of the lactonase gene from R. erythropolis and also its heterologous expression in Escherichia coli. The γ-lactone catabolic pathway is induced by pathogen communication rather than by pathogen invasion. This is thus a novel and unusual biocontrol pathway, differing from those previously described as protecting plants from phytopathogens. These findings also suggest the existence of an additional pathway contributing to plant protection.

  2. Entomopathogenic fungi as potential biocontrol agents of the ecto-parasitic mite, Varroa destructor, and their effect on the immune response of honey bees (Apis mellifera L.).

    Science.gov (United States)

    Hamiduzzaman, Mollah Md; Sinia, Alice; Guzman-Novoa, Ernesto; Goodwin, Paul H

    2012-11-01

    Three isolates of each of the entomopathogenic fungi, Metarhizium anisopliae, Beauveria bassiana and Clonostachys rosea, were assessed for their pathogenicity to the honey bee parasitic mite, Varroa destructor. The fungi were applied to varroa mites by immersing them in a spore solution, and then the inoculated mites were placed on honey bee brood inside capped cells. At 7 days post inoculation (dpi), the three fungi caused significant varroa mortality compared to non-inoculated mites. In brood treated only with varroa mites, expression of the honey bee genes, hymenoptaecin and poly U binding factor 68 Kd (pUf68), decreased over time, while expression of blue cheese (BlCh) and single minded (SiMd) was not affected. In brood inoculated directly only with M. anisopliae or B. bassiana, the emerged adults showed reduced weight indicating infection by the fungi, which was confirmed by observation of hyphae in the brood. Fungal infection of the brood resulted in increased expression of hymenoptaecin, pUf68 and BlCh, but not SiMd. In brood treated with varroa mites that had been inoculated with the fungi, expression of hymenoptaecin, pUf68 and BlCh, but not SiMd, was even more up-regulated. While varroa mites can suppress gene expression in honey bee brood, varroa mites infected with entomopathogenic fungi induced their expression. This may be due to a low level of fungal infection of the bee, which negated the immunosuppression by the mites. Therefore, entomopathogenic fungi could reduce varroa mite damage to honey bee brood by both infecting the parasite and preventing varroa-associated suppression of honey bee immunity.

  3. Identification and Quantification of a Toxigenic and Non-Toxigenic Aspergillus flavus Strain in Contaminated Maize Using Quantitative Real-Time PCR.

    Science.gov (United States)

    Mylroie, J Erik; Ozkan, Seval; Shivaji, Renuka; Windham, Gary L; Alpe, Michael N; Williams, W Paul

    2016-01-01

    Aflatoxins, which are produced by Aspergillus flavus, are toxic to humans, livestock, and pets. The value of maize (Zea mays) grain is markedly reduced when contaminated with aflatoxin. Plant resistance and biological control using non-toxin producing strains are considered effective strategies for reducing aflatoxin accumulation in maize grain. Distinguishing between the toxin and non-toxin producing strains is important in determining the effectiveness of bio-control strategies and understanding inter-strain interactions. Using polymorphisms found in the fungal rRNA intergenic spacer region (IGS) between a toxigenic strain of A. flavus (NRRL 3357) and the non-toxigenic strain used in the biological control agent Afla-Guard(®) (NRRL 21882), we developed a set of primers that allows for the identification and quantification of the two strains using quantitative PCR. This primer set has been used to screen maize grain that was inoculated with the two strains individually and co-inoculated with both strains, and it has been shown to be effective in both the identification and quantification of both strains. Screening of co-inoculated ears from multiple resistant and susceptible genotypic crosses revealed no significant differences in fungal biomass accumulation of either strain in the field tests from 2010 and 2011 when compared across the means of all genotypes. Only one genotype/year combination showed significant differences in strain accumulation. Aflatoxin accumulation analysis showed that, as expected, genotypes inoculated with the toxigenic strain accumulated more aflatoxin than when co-inoculated with both strains or inoculated with only the non-toxigenic strain. Furthermore, accumulation of toxigenic fungal mass was significantly correlated with aflatoxin accumulation while non-toxigenic fungal accumulation was not. This primer set will allow researchers to better determine how the two fungal strains compete on the maize ear and investigate the interaction

  4. Identification and Quantification of a Toxigenic and Non-Toxigenic Aspergillus flavus Strain in Contaminated Maize Using Quantitative Real-Time PCR

    Directory of Open Access Journals (Sweden)

    J. Erik Mylroie

    2016-01-01

    Full Text Available Aflatoxins, which are produced by Aspergillus flavus, are toxic to humans, livestock, and pets. The value of maize (Zea mays grain is markedly reduced when contaminated with aflatoxin. Plant resistance and biological control using non-toxin producing strains are considered effective strategies for reducing aflatoxin accumulation in maize grain. Distinguishing between the toxin and non-toxin producing strains is important in determining the effectiveness of bio-control strategies and understanding inter-strain interactions. Using polymorphisms found in the fungal rRNA intergenic spacer region (IGS between a toxigenic strain of A. flavus (NRRL 3357 and the non-toxigenic strain used in the biological control agent Afla-Guard® (NRRL 21882, we developed a set of primers that allows for the identification and quantification of the two strains using quantitative PCR. This primer set has been used to screen maize grain that was inoculated with the two strains individually and co-inoculated with both strains, and it has been shown to be effective in both the identification and quantification of both strains. Screening of co-inoculated ears from multiple resistant and susceptible genotypic crosses revealed no significant differences in fungal biomass accumulation of either strain in the field tests from 2010 and 2011 when compared across the means of all genotypes. Only one genotype/year combination showed significant differences in strain accumulation. Aflatoxin accumulation analysis showed that, as expected, genotypes inoculated with the toxigenic strain accumulated more aflatoxin than when co-inoculated with both strains or inoculated with only the non-toxigenic strain. Furthermore, accumulation of toxigenic fungal mass was significantly correlated with aflatoxin accumulation while non-toxigenic fungal accumulation was not. This primer set will allow researchers to better determine how the two fungal strains compete on the maize ear and investigate

  5. The wMel strain of Wolbachia Reduces Transmission of Zika virus by Aedes aegypti

    Science.gov (United States)

    Aliota, Matthew T.; Peinado, Stephen A.; Velez, Ivan Dario; Osorio, Jorge E.

    2016-01-01

    Zika virus (ZIKV) is causing an explosive outbreak of febrile disease in the Americas. There are no effective antiviral therapies or licensed vaccines for this virus, and mosquito control strategies have not been adequate to contain the virus. A promising candidate for arbovirus control and prevention relies on the introduction of the intracellular bacterium Wolbachia into Aedes aegypti mosquitoes. This primarily has been proposed as a tool to control dengue virus (DENV) transmission; however, evidence suggests Wolbachia infections confer protection for Ae. aegypti against other arboviruses. At present, it is unknown whether or not ZIKV can infect, disseminate, and be transmitted by Wolbachia-infected Ae. aegypti. Using Ae. aegypti infected with the wMel strain of Wolbachia that are being released in Medellin, Colombia, we report that these mosquitoes have reduced vector competence for ZIKV. These results support the use of Wolbachia biocontrol as a multivalent strategy against Ae. aegypti-transmitted viruses. PMID:27364935

  6. The wMel strain of Wolbachia Reduces Transmission of Zika virus by Aedes aegypti.

    Science.gov (United States)

    Aliota, Matthew T; Peinado, Stephen A; Velez, Ivan Dario; Osorio, Jorge E

    2016-01-01

    Zika virus (ZIKV) is causing an explosive outbreak of febrile disease in the Americas. There are no effective antiviral therapies or licensed vaccines for this virus, and mosquito control strategies have not been adequate to contain the virus. A promising candidate for arbovirus control and prevention relies on the introduction of the intracellular bacterium Wolbachia into Aedes aegypti mosquitoes. This primarily has been proposed as a tool to control dengue virus (DENV) transmission; however, evidence suggests Wolbachia infections confer protection for Ae. aegypti against other arboviruses. At present, it is unknown whether or not ZIKV can infect, disseminate, and be transmitted by Wolbachia-infected Ae. aegypti. Using Ae. aegypti infected with the wMel strain of Wolbachia that are being released in Medellin, Colombia, we report that these mosquitoes have reduced vector competence for ZIKV. These results support the use of Wolbachia biocontrol as a multivalent strategy against Ae. aegypti-transmitted viruses. PMID:27364935

  7. The Role of Chitinase Production by Stenotrophomonas maltophilia Strain C3 in Biological Control of Bipolaris sorokiniana.

    Science.gov (United States)

    Zhang, Z; Yuen, G Y

    2000-04-01

    ABSTRACT The role of chitinase production by Stenotrophomonas maltophilia strain C3 in biological control of leaf spot on tall fescue (Festuca arundinacea), caused by Bipolaris sorokiniana, was investigated in vitro and in vivo. The filtrate of a broth culture of C3, with chitin as the carbon source, was separated into fractions. A high molecular-weight fraction (>8 kDa) was chitinolytic and more inhibitory than a low-molecular-weight, nonchitinolytic fraction to conidial germination and hyphal growth by B. sorokiniana and to leaf spot development. A protein fraction derived by ammonium sulfate precipitation and a chitinase fraction purified by chitin affinity chromatography also were chitinolytic and highly antifungal. The chitinolytic fractions caused swelling and vacuolation of conidia and discoloration, malformation, and degradation of germ tubes. When boiled, the chitinolytic fractions lost chitinase activity along with most of the antifungal properties. Two chitinase-deficient and two chitinase-reduced mutants of C3 were compared with the wild-type strain for inhibition of germination of B. sorokiniana conidia on tall fescue leaves and for suppression of leaf spot development in vivo. The mutants exhibited reduced antifungal activity and biocontrol efficacy, but did not lose all biocontrol activity. An aqueous extract of leaves colonized by wild-type C3 had higher chitinase activity than that of noncolonized leaves and was inhibitory to conidial germination. The addition of chitin to leaves along with the wild-type strain increased both chitinase and antifungal activity. The chitinase activity level of extracts from leaves colonized by a chitinase-deficient mutant of C3, with and without added chitin, was no higher than the background, and the extracts lacked antifungal activity. Chitinolysis appears to be one mechanism of biological control by strain C3, and it functions in concert with other mechanisms.

  8. Improvement of Biocontrol of Damping-off and Root Rot/Wilt of Faba Bean by Salicylic Acid and Hydrogen Peroxide.

    Science.gov (United States)

    Abdel-Monaim, Montaser Fawzy

    2013-03-01

    Rhizoctonia solani, Fusarium solani, F. oxysporum, and Macrophomina phaseolina were found to be associated with root rott and wilt symptoms of faba bean plants collected from different fieldes in New Valley governorate, Egypt. All the obtained isolates were able to attack faba bean plants (cv. Giza 40) causing damping-off and root rot/wilt diseases. R. solani isolates 2 and 5, F. solani isolate 8, F. oxysporum isolate 12 and M. phaseolina isolate 14 were the more virulent ones in the pathogenicity tests. Biocontrol agents (Trichoderma viride and Bacillus megaterium) and chemical inducers (salicylic acid [SA] and hydrogen peroxide) individually or in combination were examined for biological control of damping-off and root rot/wilt and growth promoting of faba bean plants in vitro and in vivo. Both antagonistic biocontrol agents and chemical inducers either individually or in combination inhibited growth of the tested pathogenic fungi. Biocontrol agents combined with chemical inducers recorded the highest inhibited growth especially in case SA + T. viride and SA + B. megaterium. Under green house and field conditions, all treatments significantly reduced damping-off and root rot/wilt severity and increased of survival plants. Also, these treatments increased fresh and weights of the survival plants in pots compared with control. The combination between biocontrol agents and chemical inducers were more effective than used of them individually and SA + T. viride was the best treatment in this respect. Also, under field conditions, all these treatments significantly increased growth parameters (plant height and number of branches per plant) and yield components (number of pods per plant and number of seeds per plant, weight of 100 seeds and total yield per feddan) and protein content in both seasons (2010~2011 and 2011~2012). Faba bean seeds soaked in SA + T. viride and SA + B. megaterium were recorded the highest growth parameters and yield components. Generally, the

  9. Biocontrol of Aspergillus species on peanut kernels by antifungal diketopiperazine producing Bacillus cereus associated with entomopathogenic nematode.

    Directory of Open Access Journals (Sweden)

    Sasidharan Nishanth Kumar

    Full Text Available The rhabditid entomopathogenic nematode associated Bacillus cereus and the antifungal compounds produced by this bacterium were evaluated for their activity in reducing postharvest decay of peanut kernels caused by Aspergillus species in in vitro and in vivo tests. The results showed that B. cereus had a significant effect on biocontrol effectiveness in in vitro and in vivo conditions. The antifungal compounds produced by the B. cereus were purified using silica gel column chromatography and their structure was elucidated using extensive spectral analyses. The compounds were identified as diketopiperazines (DKPs [cyclo-(L-Pro-Gly, cyclo(L-Tyr-L-Tyr, cyclo-(L-Phe-Gly and cyclo(4-hydroxy-L-Pro-L-Trp]. The antifungal activities of diketopiperazines were studied against five Aspergillus species and best MIC of 2 µg/ml was recorded against A. flavus by cyclo(4-hydroxy-L-Pro-L-Trp. To investigate the potential application of cyclo(4-hydroxy-L-Pro-L-Trp to eliminate fungal spoilage in food and feed, peanut kernels was used as a food model system. White mycelia and dark/pale green spores of Aspergillus species were observed in the control peanut kernels after 2 days incubation. However the fungal growth was not observed in peanut kernels treated with cyclo(4-hydroxy-L-Pro-L-Trp. The cyclo(4-hydroxy-L-Pro-L-Trp was nontoxic to two normal cell lines [fore skin (FS normal fibroblast and African green monkey kidney (VERO] up to 200 µg/ml in MTT assay. Thus the cyclo(4-hydroxy-L-Pro-L-Trp identified in this study may be a promising alternative to chemical preservatives as a potential biopreservative agent which prevent fungal growth in food and feed. To the best of our knowledge, this is the first report demonstrating that the entomopathogenic nematode associated B. cereus and cyclo(4-hydroxy-L-Pro-L-Trp could be used as a biocontrol agents against postharvest fungal disease caused by Aspergillus species.

  10. Strains and Sprains

    Science.gov (United States)

    ... ON THIS TOPIC Sports Medicine Center Preventing Children's Sports Injuries Computer-Related Repetitive Stress Injuries Knee Injuries Broken ... Knee Injuries Sports and Exercise Safety Dealing With Sports Injuries Groin Strain Ankle Sprains Strains and Sprains Stretching ...

  11. Strained Silicon Photonics

    OpenAIRE

    Wehrspohn, Ralf B; Jörg Schilling; Christian Bohley; Clemens Schriever

    2012-01-01

    A review of recent progress in the field of strained silicon photonics is presented. The application of strain to waveguide and photonic crystal structures can be used to alter the linear and nonlinear optical properties of these devices. Here, methods for the fabrication of strained devices are summarized and recent examples of linear and nonlinear optical devices are discussed. Furthermore, the relation between strain and the enhancement of the second order nonlinear susceptibility is inves...

  12. Endophytic fungus strain 28 isolated from Houttuynia cordata possesses wide-spectrum antifungal activity.

    Science.gov (United States)

    Pan, Feng; Liu, Zheng-Qiong; Chen, Que; Xu, Ying-Wen; Hou, Kai; Wu, Wei

    2016-01-01

    The aim of this paper is to identify and investigate an endophytic fungus (strain 28) that was isolated from Houttuynia cordata Thunb, a famous and widely-used Traditional Chinese Medicine. Based on morphological methods and a phylogenetic analysis of ITS sequences, this strain was identified as Chaetomium globosum. An antifungal activity bioassay demonstrated that the crude ethyl acetate (EtOAc) extracts of strain 28 had a wide antifungal spectrum and strong antimicrobial activity, particularly against Exserohilum turcicum (Pass.) Leonard et Suggs, Botrytis cinerea persoon and Botrytis cinerea Pers. ex Fr. Furthermore, the fermentation conditions, extraction method and the heat stability of antifungal substances from strain 28 were also studied. The results showed that optimal antifungal activity can be obtained with the following parameters: using potato dextrose broth (PDB) as the base culture medium, fermentation for 4-8d (initial pH: 7.5), followed by extraction with EtOAc. The extract was stable at temperatures up to 80°C. This is the first report on the isolation of endophytic C. globosum from H. cordata to identify potential alternative biocontrol agents that could provide new opportunities for practical applications involving H. cordata. PMID:26991297

  13. Induced systemic resistance against Botrytis cinerea by Micromonospora strains isolated from root nodules

    Directory of Open Access Journals (Sweden)

    Pilar eMartínez-Hidalgo

    2015-09-01

    Full Text Available Micromonospora is a Gram positive bacterium that can be isolated from nitrogen fixing nodules from healthy leguminous plants, where they could be beneficial to the plant. Their plant growth promoting activity in legume and non-legume plants has been previously demonstrated. The present study explores the ability of Micromonospora strains to control fungal pathogens and to stimulate plant immunity. Micromonospora strains isolated from surface sterilized nodules of alfalfa showed in vitro antifungal activity against several pathogenic fungi. Moreover, root inoculation of tomato plants with these Micromonospora strains effectively reduced leaf infection by the fungal pathogen Botrytis cinerea, despite spatial separation between both microorganisms. This induced systemic resistance, confirmed in different tomato cultivars, is long lasting. Gene expression analyses evidenced that Micromonospora stimulates the plant capacity to activate defense mechanisms upon pathogen attack. The defensive response of tomato plants inoculated with Micromonospora spp. differs from that of non-inoculated plants, showing a stronger induction of jasmonate-regulated defenses when the plant is challenged with a pathogen. The hypothesis of jasmonates playing a key role in this defense priming effect was confirmed using defense-impaired tomato mutants, since the JA-deficient line def1 was unable to display a long term induced resistance upon Micromonospora spp. inoculation.In conclusion, nodule isolated Micromonospora strains should be considered excellent candidates as biocontrol agents as they combine both direct antifungal activity against plant pathogens and the ability to prime plant immunity.

  14. Induced systemic resistance against Botrytis cinerea by Micromonospora strains isolated from root nodules.

    Science.gov (United States)

    Martínez-Hidalgo, Pilar; García, Juan M; Pozo, María J

    2015-01-01

    Micromonospora is a Gram positive bacterium that can be isolated from nitrogen fixing nodules from healthy leguminous plants, where they could be beneficial to the plant. Their plant growth promoting activity in legume and non-legume plants has been previously demonstrated. The present study explores the ability of Micromonospora strains to control fungal pathogens and to stimulate plant immunity. Micromonospora strains isolated from surface sterilized nodules of alfalfa showed in vitro antifungal activity against several pathogenic fungi. Moreover, root inoculation of tomato plants with these Micromonospora strains effectively reduced leaf infection by the fungal pathogen Botrytis cinerea, despite spatial separation between both microorganisms. This induced systemic resistance, confirmed in different tomato cultivars, is long lasting. Gene expression analyses evidenced that Micromonospora stimulates the plant capacity to activate defense mechanisms upon pathogen attack. The defensive response of tomato plants inoculated with Micromonospora spp. differs from that of non-inoculated plants, showing a stronger induction of jasmonate-regulated defenses when the plant is challenged with a pathogen. The hypothesis of jasmonates playing a key role in this defense priming effect was confirmed using defense-impaired tomato mutants, since the JA-deficient line def1 was unable to display a long term induced resistance upon Micromonospora spp. inoculation. In conclusion, nodule isolated Micromonospora strains should be considered excellent candidates as biocontrol agents as they combine both direct antifungal activity against plant pathogens and the ability to prime plant immunity. PMID:26388861

  15. Pyrrolnitrin and Hydrogen Cyanide Production by Pseudomonas chlororaphis Strain PA23 Exhibits Nematicidal and Repellent Activity against Caenorhabditis elegans.

    Science.gov (United States)

    Nandi, Munmun; Selin, Carrie; Brassinga, Ann Karen C; Belmonte, Mark F; Fernando, W G Dilantha; Loewen, Peter C; de Kievit, Teresa R

    2015-01-01

    Pseudomonas chlororaphis strain PA23 is a biocontrol agent able to suppress growth of the fungal pathogen Sclerotinia sclerotiorum. This bacterium produces an arsenal of exometabolites including pyrrolnitrin (PRN), phenazine (PHZ), hydrogen cyanide (HCN), and degradative enzymes. Production of these compounds is controlled at both the transcriptional and posttranscriptional levels by the Gac-Rsm system, RpoS, PsrA, and the Phz quorum-sensing system. Beyond pathogen-suppression, the success of a biocontrol agent is dependent upon its ability to establish itself in the environment where predation by bacterivorous organisms, including nematodes, may threaten persistence. The focus of this study was to investigate whether PA23 is able to resist grazing by Caenorhabditis elegans and to define the role played by exoproducts in the bacterial-nematode interaction. We discovered that both PRN and HCN contribute to fast- and slow-killing of C. elegans. HCN is well-established as having lethal effects on C. elegans; however, PRN has not been reported to be nematicidal. Exposure of L4 stage nematodes to purified PRN reduced nematode viability in a dose-dependent fashion and led to reduced hatching of eggs laid by gravid adults. Because bacterial metabolites can act as chemoattractants or repellents, we analyzed whether PA23 exhibited attractant or repulsive properties towards C. elegans. Both PRN and HCN were found to be potent repellents. Next we investigated whether the presence of C. elegans would elicit changes in PA23 gene activity. Co-culturing the two organisms increased expression of a number of genes associated with biocontrol, including phzA, hcnA, phzR, phzI, rpoS and gacS. Exoproduct analysis showed that PHZ and autoinducer signals were upregulated, consistent with the gene expression profiles. Collectively, these findings indicate that PA23 is able to sense the presence of C. elegans and it is able to both repel and kill the nematodes, which should facilitate

  16. SSR Markers for Trichoderma virens: Their Evaluation and Application to Identify and Quantify Root-Endophytic Strains

    Directory of Open Access Journals (Sweden)

    Joerg Geistlinger

    2015-11-01

    Full Text Available Using biological fertilizers and pesticides based on beneficial soil microbes in order to reduce mineral fertilizers and chemical pesticides in conventional agriculture is still a matter of debate. In this regard, a European research project seeks to elucidate the role of root-endophytic fungi and to develop molecular tools to trace and quantify these fungi in the rhizosphere and root tissue. To do this, the draft genome sequence of the biocontrol fungus Trichoderma virens (T. virens was screened for simple sequence repeats (SSRs and primers were developed for 12 distinct loci. Primers were evaluated using a global collection of ten isolates where an average of 7.42 alleles per locus was detected. Nei’s standard genetic distance ranged from 0.18 to 0.27 among the isolates, and the grand mean of haploid diversity in AMOVA analysis was 0.693 ± 0.019. Roots of tomato plants were inoculated with different strains and harvested six weeks later. Subsequent PCR amplification identified root-endophytic strains and co-colonization of roots by different strains. Markers were applied to qPCR to quantify T. virens strains in root tissue and to determine their identity using allele-specific melting curve analysis. Thus, the root-endophytic lifestyle of T. virens was confirmed, strains in roots were quantified and simultaneous colonization of roots by different strains was observed.

  17. Isolation and Selection of Epiphytic Yeast for Biocontrol of Botrytis cinerea Pers. on Table Grapes Aislación y Selección de Levaduras Epífitas para el Biocontrol de Botrytis cinerea Pers. en Uva de Mesa

    Directory of Open Access Journals (Sweden)

    Marisol Vargas

    2012-09-01

    Full Text Available Botrytis cinerea Pers., the causal agent of gray mold, infects more than 200 plant species. This pathogen has traditionally been controlled by fungicides. However, with the increasing demand for pesticide-free foods new control strategies are needed. The objective of this study was to isolate and select grapevine (Vitis vinifera L. epiphytic yeasts for the biocontrol of B. cinerea in table grapes. Of the total isolated yeasts (n = 256, 32 exhibited mycelial growth inhibition in dual cultures with a halo > 4 mm, and eight of these isolates inhibited > 90% of conidial germination. When evaluating increasing concentrations on conidial germination inhibition, a dose-dependent response was observed with EC90 values from 0.45 x 10(5 to 0.22 x 10(8 cells mL-1. The antagonistic activity of six yeasts against B. cinerea in table grape berries 'Flame Seedless' increased as the yeast colonization time increased from 1 to 24 h on the berries, resulting in a higher biocontrol activity on B. cinerea. These results show the effectiveness of grapevine epiphytic yeasts as biocontrol agents of B. cinerea on table grapes.Botrytis cinerea Pers., agente causal de la pudrición gris, infecta a más de 200 especies vegetales. Tradicionalmente, este patógeno ha sido controlado con fungicidas; sin embargo, la creciente demanda de alimentos libres de pesticidas hace necesario el uso de nuevas estrategias de control. El objetivo de este estudio fue aislar y seleccionar levaduras epífitas de vid (Vitis vinifera L. para el biocontrol de B. cinerea en uva de mesa. Del total de levaduras aisladas (n = 256, 32 presentaron inhibición del crecimiento micelial, en cultivos duales, con un halo > 4 mm y ocho de estos aislamientos inhibieron la germinación de conidias > 90%. Al evaluar concentraciones crecientes de levaduras sobre la inhibición de la germinación de conidias, se observó una respuesta dosis-dependiente, con valores de CE90 de 0,45 x 10(5 a 0,22 x 10(8 c

  18. Biocontrol of postharvest gray and blue mold decay of apples with Rhodotorula mucilaginosa and possible mechanisms of action.

    Science.gov (United States)

    Li, Renping; Zhang, Hongyin; Liu, Weimin; Zheng, Xiaodong

    2011-03-30

    The efficacy of Rhodotorula mucilaginosa against postharvest gray mold, blue mold and natural decay development of apples and the possible mechanisms involved were investigated. The decay incidence and lesion diameter of gray mold and blue mold of apples treated by R. mucilaginosa were significantly reduced compared with the control fruits, and the higher concentration of R. mucilaginosa, the better the efficacy of the biocontrol. R. mucilaginosa also significantly reduced the natural decay development of apples following storage at 20°C for 35 days or at 4°C for 45 days followed by 20°C for 15 days. Germination and survival of spores of Penicillium expansum and Botrytis cinerea were markedly inhibited by R. mucilaginosa in an in vitro test. Rapid colonization of the yeast in apple wounds was observed whether stored at 20°C or 4°C. In apples, the activities of peroxidase (POD) and polyphenoloxidase (PPO) were significantly induced and lipid peroxidation (malondialdehyde (MDA) content) was highly inhibited by R. mucilaginosa treatment compared with those of the control fruits. All these results indicated that R. mucilaginosa has great potential for development of commercial formulations to control postharvest pathogens on fruits. Its modes of action were based on competition for space and nutrients with pathogens, inducement of activities of defense-related enzymes such as POD, PPO and inhibition of lipid peroxidation (MDA content) of apples, so as to enhance the resistance and delay the ripening and senescence of apples.

  19. Conservation of bio-control agents in cotton, gossypium hirsutum l. field by food supplements for insect pests management

    International Nuclear Information System (INIS)

    The study reports the use of artificial food sprays to conserve the parasitoids and predators for the management of insect pests in cotton field. Cotton crop was treated with bio-control agents, Chrysoperla carnea and Trichogramma chilonis alongwith different food attractants such as Protein hydrolysate and sugar alone and in combination in a randomized complete block design. Each treatment was applied on one-acre field with three replications. Results showed that the chemicals tested helped in increasing the populations of beneficial insects including; C. carnea, T. chilonis and Orius spp., in the field. The populations of C. carnea and T. chilonis were found the highest in the combined treatment of protein hydrolysate and sugar as compared to other treatments where protein hydrolysate and sugar were used separately. However, the population of Orius spp. was higher in the treatment where only sugar solution was sprayed as food supplement. Consequently, incorporation of food supplements in the trial increased the establishment of natural enemies and subsequently the predation/ parasitism percentage enhanced on the insect pests of cotton. (author)

  20. Effects of local vegetation and plantation age for the parasitoid Asecodes mento-a biocontrol agent in organic strawberry fields

    Institute of Scientific and Technical Information of China (English)

    Johan A.Stenberg

    2012-01-01

    The parasitoid Asecodes mento (Walker,1839) (Hymenoptera:Eulophidae)is the most important biocontrol agent of the strawberry leaf beetle Galerucella tenella (L.) (Coleoptera:Chrysomelidae) in northern Europe.Here,I investigated whether natural parasitism in organic strawberry plantations was affected by the presence of the alternative host plant meadowsweet (Filipendula ulmaria),and whether parasitism rates differed between plantations of different ages (6 to 79 years).I also investigated whether parasitoid brood size,body size and sex ratio differed between the two host plants in the field.Parasitism was very low (0%) in newly established plantations and increased to a plateau (~40%) in fields where strawberries had been grown for approximately 20 years or longer.Such an extended colonization process is unacceptable for commercial growers.It would thus be desirable to find a method to catalyze parasitoid population buildup in young plantations.Parasitoid brood sizes were larger in beetles collected from meadowsweet,while body size and sex ratio did not differ between parasitoids collected from the two plants.These findings suggest that meadowsweet can export parasitoids to neighboring strawberry fields.Although this is a possibility,I did not find any significant differences in parasitism rates between isolated strawberry fields and fields adjacent to meadowsweet stands,indicating that effects of local vegetation are small on parasitism rates.Releasing parasitoids in newly established strawberry plantations may be a better strategy for quickly obtaining high parasitism than intercropping with meadowsweet.

  1. Quantifying the associations between fungal endophytes and biocontrol-induced herbivory of invasive purple loosestrife (Lythrum salicaria L.).

    Science.gov (United States)

    David, Aaron S; Quiram, Gina L; Sirota, Jennie I; Seabloom, Eric W

    2016-01-01

    Fungal endophytes are one of several groups of heterotrophic organisms that associate with living plants. The net effects of these groups of organisms on each other and ultimately on their host plants depend in part on how they facilitate or antagonize one another. In this study we quantified the associations between endophyte communities and herbivory induced by a biological control in the invasive Lythrum salicaria at various spatial scales using a culture-based approach. We found positive associations between herbivory damage and endophyte isolation frequency and richness at the site level and weak, positive associations at the leaf level. Herbivory damage was more strongly influenced by processes at the site level than were endophyte isolation frequency and community structure, which were influenced by processes at the plant and leaf levels. Furthermore, endophytic taxa found in low herbivory sites were nested subsets of those taxa found at high herbivory sites. Our findings suggest that endophyte communities of L. salicaria are associated with, and potentially facilitated by, biocontrol-induced herbivory. Quantifying the associations between heterotrophic groups ultimately may lead to a clearer understanding of their complex interactions with plants. PMID:27091387

  2. Biology and host preferences of Cryptorhynchus melastomae (Coleoptera: Curculionidae), a possible biocontrol agent for Miconia calvescens (Melastomataceae) in Hawaii.

    Science.gov (United States)

    Reichert, Elisabeth; Johnson, M Tracy; Chacón, Eduardo; Anderson, Robert S; Wheeler, Terry A

    2010-12-01

    The introduced plant Miconia calvescens (Melastomataceae) poses a grave threat to Hawaii's native ecosystems and biodiversity. One potential candidate for classical biological control is Cryptorhynchus melastomae (Coleoptera: Curculionidae: Cryptorhynchinae), a stem-boring weevil from Central and South America. This weevil feeds on M. calvescens in its native Costa Rica and has been successfully reared under greenhouse conditions. Comparison of its environmental conditions in Costa Rica with those in the Miconia infested areas of Hawaii indicates the latter is a suitable habitat for C. melastomae. C. melastomae has one or two generations per year. Adults feed on new stems, petioles, leaf buds, veins, and lamina, whereas larvae mine the stem until pupation. Adults appear to prefer saplings for oviposition and feeding. Under greenhouse conditions both adults and larvae can seriously damage and kill small M. calvescens. Preliminary host testing indicates that C. melastomae may be family specific on Melastomataceae. However, because Hawaii lacks native melastomes and has many other serious melastome weeds, a family specific insect may be suitable as a biocontrol agent in this case. PMID:22182550

  3. Biocontrol agents-mediated suppression of oxalic acid induced cell death during Sclerotinia sclerotiorum-pea interaction.

    Science.gov (United States)

    Jain, Akansha; Singh, Akanksha; Singh, Surendra; Sarma, Birinchi Kumar; Singh, Harikesh Bahadur

    2015-05-01

    Oxalic acid (OA) is an important pathogenic factor during early Sclerotinia sclerotiorum-host interaction and might work by reducing hydrogen peroxide production (H2 O2 ). In the present investigation, oxalic acid-induced cell death in pea was studied. Pea plants treated with biocontrol agents (BCAs) viz., Pseudomonas aeruginosa PJHU15, Bacillus subtilis BHHU100, and Trichoderma harzianum TNHU27 either singly and/or in consortium acted on S. sclerotiorum indirectly by enabling plants to inhibit the OA-mediated suppression of oxidative burst via induction of H2 O2 . Our results showed that BCA treated plants upon treatment with culture filtrate of the pathogen, conferred the resistance via. significantly decreasing relative cell death of pea against S. sclerotiorum compared to control plants without BCA treatment but treated with the culture filtrate of the pathogen. The results obtained from the present study indicate that the microbes especially in consortia play significant role in protection against S. sclerotiorum by modulating oxidative burst and partially enhancing tolerance by increasing the H2 O2 generation, which is otherwise suppressed by OA produced by the pathogen.

  4. In vitro evaluation of Pseudomonas bacterial isolates from rice phylloplane for biocontrol of Rhizoctonia solani and plant growth promoting traits.

    Science.gov (United States)

    Akter, Shamima; Kadir, Jugah; Juraimi, Abdul Shukor; Saud, Halimi Mohd

    2016-07-01

    The ability for biocontrol and plant growth promotion of three Pseudomonas bacterial isolates namely Pseudomonas fluorescens (UMB20), Pseudomonas aeruginosa (KMB25) and Pseudomonas asplenii (BMB42) obtained from rice plants was investigated. Fungal growth inhibition by the isolates ranged from 86.85 to 93.15% in volatile and 100% in diffusible metabolites test. Among the isolates, BMB42 showed fungal growth inhibition significantly in the volatile metabolite test. Isolates UMB20 and BMB42 were able to synthesis chitinase with chitinolytic indices of 13.66 and 13.50, respectively. In case of -1,3-glucanase, all the isolates showed activity to produce this enzyme at varied levels and isolate KMB25 showed significantly highest activity (53.53 ppm). Among the three isolates, KMB25 showed positive response to protease production and all of them were negative to pectinase and lipase and positive to the production of siderophore, and HCN, and were able to solubilize tricalcium phosphate. All the three bacterial isolates were capable of forming biofilm at different levels. Above results suggest that phylloplane Pseudomonas bacterial isolates have potential for antifungal activities and plant growth promotion. PMID:27498507

  5. Evaluation of biocontrol ability of native strains of Trichoderma spp on Rhizoctonia and Fusarium sp in coffee (Coffea arabica in experimental conditions

    Directory of Open Access Journals (Sweden)

    Nina Rudy

    2016-06-01

    Full Text Available Due to the indiscriminate use agrochemicals in conventional agriculture, it is causing pollution problems in the environment (soil, air and water, hence the search for alternatives that contribute to agricultural production by agro-chemical free sustainable production. This paper studies the biological control of damping off in coffee (Coffea arabica by applying antagonistic fungus Trichoderma sp. Under experimental conditions at laboratory facilities of the Academic Unit Carmen Pampa Campesina, a community of Carmen Pampa, Township Coroico. The aim of this study was to biologically control the "damping off", they found two genera that cause damping off in seedbed of coffee: Rhizoctonia sp. and Fusarium sp.To determine the percentage of growth and control in the culture medium, we used the method of counting quarters, where they gave the mycelial growth of antagonistic fungus Trichoderma sp., And the fungal pathogens Rhizoctonia sp. and Fusarium sp. Statistically there was a highly significant difference in the variable growth rate of Trichoderma sp. on pathogenic fungi Rhizoctonia sp. and Fusarium sp. at 3, 6 and 9 days that announces the time factor and treatments are interdependent. The control variable showed a highly significant difference in the time factor and treatment, but the interaction shows no significant difference this makes known factors that are independent, so the fungus Trichoderma sp. not depend on time in treatment, thus showing its inhibitory power to Rhizoctonia sp. and Fusarium sp .. This test gives references that there is antagonistic fungus control on the fungal pathogens Rhizoctonia sp. and Fusarium sp.

  6. Evaluation of biocontrol ability of native strains of Trichoderma spp on Rhizoctonia and Fusarium sp in coffee (Coffea arabica) in experimental conditions

    OpenAIRE

    Nina Rudy; Smeltekop Hugh; Almanza JC; Loza-Murguia Manuel

    2016-01-01

    Due to the indiscriminate use agrochemicals in conventional agriculture, it is causing pollution problems in the environment (soil, air and water), hence the search for alternatives that contribute to agricultural production by agro-chemical free sustainable production. This paper studies the biological control of damping off in coffee (Coffea arabica) by applying antagonistic fungus Trichoderma sp. Under experimental conditions at laboratory facilities of the Academic Unit Carmen Pampa Campe...

  7. 大豆根腐病生防菌株的筛选及鉴定%Screening and identification of biocontrol strains against soybean root rot pathogens

    Institute of Scientific and Technical Information of China (English)

    黄珊珊; 韩雪; 李丽珺; 文景芝

    2008-01-01

    采用常规方法从大豆根际分离筛选出105株对大豆根腐病菌有拮抗作用的细菌和放线菌,利用磷脂脂肪酸法和16S rDNA测序鉴定出13株细菌,分别为1株Pseudomonas putida biotype B,1株Pseudomonas chlororaphis,1株Pseudomonas sp.,4株Bacillus subtilis,2株Bacillus sp.,1株Bacillus pumilus,1株Bacillus atophaeus,1株Virgibacillus pantothenticus,1株Pantoea agglomerans GC subgroup A;10株放线菌,分别为3株Streptomyces violaceusniger violaceusniger,1株Streptoverticillium cinnamoneum,2株Streptomyces halstedii olivaceus,1株Streptomyces exfoliatus,2株Streptomyces halstedii scabies,1株Streptomyces violaceusniger hygroscopicus ossam.

  8. ISOLATION OF ENDOPHYTIC ACTINOMYCETES FROM MEDICINAL PLANTS AND ITS MUTATIONAL EFFECT IN BIOCONTROL ACTIVITY

    Directory of Open Access Journals (Sweden)

    Hema Shenpagam N.*, D. Kanchana Devi ** and Sinduja G.

    2012-11-01

    Full Text Available In the present study, the endophytic actinomycetes were collected from three medicinal plants Azadiracta indica, Ocimum sanctum and Phyllanthus amarus. Endophytic actinomycetes were isolated using different media like Starch casein agar, Starch casein nitrate agar, Actinomycetes isolation agar and Soyabean agar, while it showed more colonies in Starch casein agar. The endophytic actinomycetes were stained and biochemical tests were performed. Antimicrobial compound was purified from the filtrate by ethanol extraction method. Antagonistic activities of endophytic actinomycetes isolates were tested against bacterial pathogens (Staphylococcus aureus, Streptococcus pyogenes, Klebsiella pneumoniae and Pseudomonas aeruginosa and the fungi Rhizopus. For the selected isolates antibiotic resistance was checked using various antibiotic discs like Amoxycillin, Penicillin, Rifampicin and Ampicillin. The strains which showed efficient antibacterial activity were selected to study the effect of mutation by physical and chemical method. In this study, UV mutated endophytic actinomycetes increase antibiotic production than non-mutated endophytic Actinomycetes, whereas in chemical mutation it does not increase the antibiotic production.

  9. Characterisation of two quorum sensing systems in the endophytic Serratia plymuthica strain G3: differential control of motility and biofilm formation according to life-style

    Directory of Open Access Journals (Sweden)

    Li Jun

    2011-02-01

    Full Text Available Abstract Background N-acylhomoserine lactone (AHL-based quorum sensing (QS systems have been described in many plant-associated Gram-negative bacteria to control certain beneficial phenotypic traits, such as production of biocontrol factors and plant growth promotion. However, the role of AHL-mediated signalling in the endophytic strains of plant-associated Serratia is still poorly understood. An endophytic Serratia sp. G3 with biocontrol potential and high levels of AHL signal production was isolated from the stems of wheat and the role of QS in this isolate was determined. Results Strain G3 classified as Serratia plymuthica based on 16S rRNA was subjected to phylogenetic analysis. Using primers to conserved sequences of luxIR homologues from the Serratia genus, splIR and spsIR from the chromosome of strain G3 were cloned and sequenced. AHL profiles from strain G3 and Escherichia coli DH5α expressing splI or spsI from recombinant plasmids were identified by liquid chromatography-tandem mass spectrometry. This revealed that the most abundant AHL signals produced by SplI in E. coli were N-3-oxo-hexanoylhomoserine lactone (3-oxo-C6-HSL, N-3-oxo-heptanoylhomoserine lactone (3-oxo-C7-HSL, N-3-hydroxy-hexanoylhomoserine lactone (3-hydroxy-C6-HSL, N-hexanoylhomoserine lactone (C6-HSL, and N-heptanoyl homoserine lactone (C7-HSL; whereas SpsI was primarily responsible for the synthesis of N-butyrylhomoserine lactone (C4-HSL and N-pentanoylhomoserine lactone (C5-HSL. Furthermore, a quorum quenching analysis by heterologous expression of the Bacillus A24 AiiA lactonase in strain G3 enabled the identification of the AHL-regulated biocontrol-related traits. Depletion of AHLs with this lactonase resulted in altered adhesion and biofilm formation using a microtiter plate assay and flow cells coupled with confocal laser scanning microscopy respectively. This was different from the closely related S. plymuthica strains HRO-C48 and RVH1, where biofilm formation

  10. Selection of a Streptomyces strain able to produce cell wall degrading enzymes and active against Sclerotinia sclerotiorum.

    Science.gov (United States)

    Fróes, Adriana; Macrae, Andrew; Rosa, Juliana; Franco, Marcella; Souza, Rodrigo; Soares, Rosângela; Coelho, Rosalie

    2012-10-01

    Control of plant pathogen Sclerotinia sclerotiorum is an ongoing challenge because of its wide host range and the persistence of its sclerotia in soil. Fungicides are the most commonly used method to control this fungus but these can have ecotoxicity impacts. Chitinolytic Streptomyces strains isolated from Brazilian tropical soils were capable of inhibiting S. sclerotiorum growth in vitro, offering new possibilities for integrated pest management and biocontrol, with a new approach to dealing with an old problem. Strain Streptomyces sp. 80 was capable of irreversibly inhibiting fungal growth. Compared to other strains, its crude enzymes had the highest chitinolytic levels when measured at 25°C and strongly inhibited sclerotia from S. sclerotiorum. It produced four hydrolytic enzymes involved in fungal cell wall degradation when cultured in presence of the fungal mycelium. The best production, obtained after three days, was 0.75 U/ml for exochitinase, 0.9 U/ml for endochitinase, 0.16 U/ml for glucanase, and 1.78 U/ml for peptidase. Zymogram analysis confirmed two hydrolytic bands of chitinolytic activity with apparent molecular masses of 45.8 and 206.8 kDa. One glucanase activity with an apparent molecular mass of 55 kDa was also recorded, as well as seven bands of peptidase activity with apparent molecular masses ranging from 15.5 to 108.4 kDa. Differential interference contrast microscopy also showed alterations of hyphal morphology after co-culture. Streptomyces sp. 80 seems to be promising as a biocontrol agent against S. sclerotiorum, contributing to the development of new methods for controlling plant diseases and reducing the negative impact of using fungicides. PMID:23124748

  11. Biocide effects of volatile organic compounds produced by potential biocontrol rhizobacteria on Sclerotinia sclerotiorum

    Directory of Open Access Journals (Sweden)

    Annalisa eGiorgio

    2015-10-01

    Full Text Available Six rhizobacteria isolated from common bean and able to protect bean plants from the common bacterial blight causal agent, were in vitro evaluated for their potential antifungal effects toward different plant pathogenic fungi, mostly soil-borne. By dual culture assays, the above bacteria resulted producing diffusible and volatile metabolites which inhibited the growth of the majority of the pathogens under study. In particular, the latter substances highly affected the mycelium growth of Sclerotinia sclerotiorum strains, one of which was selected for further studies either on mycelium or sclerotia.Gas chromatographic analysis of the bacterial volatiles led to the identification of an array of volatile organic compounds (VOCs. Time course studies showed the modification of the VOCs profile along a period of 5 days. In order to evaluate the single detected VOC effects on fungal growth, some of the pure compounds were tested on S. sclerotiorum mycelium and their minimal inhibitory quantities were determined. Similarly, the minimal inhibitory quantities on sclerotia germination were also defined. Moreover, observations by light and transmission electron microscopes highlighted hyphae cytoplasm granulation and ultrastructural alterations at cell organelles, mostly membranes, mitochondria and endoplasmic reticulum. The membranes appeared one of the primary targets of bacterial volatiles, as confirmed by haemolytic activity observed for the majority of pure VOCs. However, of interest is the alteration observed on mitochondria as well.

  12. Biocide effects of volatile organic compounds produced by potential biocontrol rhizobacteria on Sclerotinia sclerotiorum.

    Science.gov (United States)

    Giorgio, Annalisa; De Stradis, Angelo; Lo Cantore, Pietro; Iacobellis, Nicola S

    2015-01-01

    Six rhizobacteria isolated from common bean and able to protect bean plants from the common bacterial blight (CBB) causal agent, were in vitro evaluated for their potential antifungal effects toward different plant pathogenic fungi, mostly soil-borne. By dual culture assays, the above bacteria resulted producing diffusible and volatile metabolites which inhibited the growth of the majority of the pathogens under study. In particular, the latter substances highly affected the mycelium growth of Sclerotinia sclerotiorum strains, one of which was selected for further studies either on mycelium or sclerotia. Gas chromatographic analysis of the bacterial volatiles led to the identification of an array of volatile organic compounds (VOCs). Time course studies showed the modification of the VOCs profile along a period of 5 days. In order to evaluate the single detected VOC effects on fungal growth, some of the pure compounds were tested on S. sclerotiorum mycelium and their minimal inhibitory quantities were determined. Similarly, the minimal inhibitory quantities on sclerotia germination were also defined. Moreover, observations by light and transmission electron microscopes highlighted hyphae cytoplasm granulation and ultrastructural alterations at cell organelles, mostly membranes, mitochondria, and endoplasmic reticulum. The membranes appeared one of the primary targets of bacterial volatiles, as confirmed by hemolytic activity observed for the majority of pure VOCs. However, of interest is the alteration observed on mitochondria as well. PMID:26500617

  13. Isolation and evaluation of biocontrol agents in controlling anthracnose disease of mango in Thailand

    Directory of Open Access Journals (Sweden)

    Rungjindamai Nattawut

    2016-07-01

    Full Text Available The agricultural based economy is a core business in Thailand and food export is one of the main sources of income for the Thai population. However, pesticides are overused and misused. As a result there is an urgent need to reduce the use of synthetic chemicals. Biological control offers an alternative to the use of pesticides. Mango (Mangifera indica L. is widely planted in Thailand and is one of the major cash crops for international export. However, mango suffers from various diseases especially anthracnose, a fungal disease caused by Colletotrichum gloeosporioides. One hundred and twelve isolates of epiphytic microbes were isolated from healthy leaves and fruits of mangoes; this included 93 and 19 isolates of epiphytic bacteria and yeasts, respectively. They were screened for bioactivity against a pathogenic strain of C. gloeosporioides isolated from diseased mangoes using a dual culture technique. Out of 112 isolates, eight isolates exhibited at least 60% inhibition. These isolates were further screened for their inhibition on mango using fruit inoculation. Two isolates reduced the lesion sizes caused by C. gloeosporioides compared to control treatment. These two isolates, based on phenotypical and biochemical tests, were identified as Bacillus sp. MB61 and Bacillus sp. LB72.

  14. Osmotically induced trehalose and glycine betaine accumulation improves tolerance to desiccation, survival and efficacy of the postharvest biocontrol agent Pantoea agglomerans EPS125.

    Science.gov (United States)

    Bonaterra, Anna; Camps, Jaume; Montesinos, Emilio

    2005-09-01

    The application of the biocontrol agent Pantoea agglomerans EPS125 to unwounded fruits was practically ineffective for control of postharvest blue mould caused by Penicillium expansum when the treatment and subsequent wounding and pathogen inoculation were separated by periods of unfavourable conditions. This was due to a rapid decrease in viability of the alocthonous introduced biocontrol agent in the intact peel surface. A system for osmoadaptation of the biocontrol agent was developed by combining saline osmotic stress and osmolyte amendment to the growth medium. Osmoadapted cells accumulated trehalose and glycine betaine (GB) intracellularly and showed a higher tolerance to desiccation than non-osmoadapted cells. Osmoadaptation in NaCl plus GB during inoculum preparation increased considerably survival on the peel surface of apple fruits. This effect was significant under low relative humidity (RH) and fluctuating RH conditions, but was not significant at high RH. Osmoadaptation significantly improved blue mould control under conditions where the standard biological control treatments were ineffective. The rot diameter was significantly reduced in apple fruits which were treated with EPS125 and incubated for several days under low, high or fluctuating RH, followed by wounding and inoculation of P. expansum. Growth of EPS125 with NaCl, either with or without the addition of GB, was an effective osmoadaptation treatment for improving blue mould rot control. However, the addition of GB to the NaCl amended growth medium increased 4-5-fold growth rate and OD of the cultures. This is an advantage for mass production of P. agglomerans EPS125 in a NaCl amended growth medium.

  15. 保护地土壤生防木霉菌种群多样性研究%Study on Genetic Diversity of Biocontrol Trichoderma Population in Greenhouse Soils

    Institute of Scientific and Technical Information of China (English)

    贺字典; 高增贵; 高玉峰; 张洋

    2011-01-01

    采用传统的形态学特征和分子方法(ITS、TEF序列和UP-PCR)研究了蔬菜保护地土壤中木霉菌种群多样性及其影响因素.结果表明:提交genbank 11个木霉种的序列,分别为长枝木霉(Trichoderma longibrachiatum)、拟康氏木霉(Trichoderma pseudokoningii)、黄绿木霉(Trichoderma aureoviride)、棘孢木霉(Trichoderma asperellum)、深绿木霉(Trichoderma atroviride)、哈茨木霉(Trichoderma harzianum)、非钩木霉(Trichoderma inhamatum)、微孢木霉(Trichoderma minutisporum)、长孢木霉(Trichoderma longipile)和粘绿木霉(Trichoderma virens);木霉菌24个菌株经UP-PCR扩增,引物AS4、AS19、L45扩增出一条500 bp大小的木霉菌种的特征性谱带,其他谱带则为多态性谱带,多态性达93.5%;营养条件、杀菌剂及土壤因素对不同种木霉菌的影响不同,得到2株适应性较强的木霉菌株,有望成为生防菌株.%11 Trichoderma species were submitted to genbank by ITS ,TEF sequences analysis methods and effect factors of population diversity were studied in this paper, which included T. Longibrachiatum, T. Atroviride, T. Harziamun, T. Viren, T. Minutisporum, T. Pseudokoningii, T. Aureoviride, T. Inhamatum, T. Asperellum, T. Longipile and T. Helicum. At the same time, genetic diversity of Tnchoderma isolates were analyzed by Universally Primed PCR. A feature band ,500 bp, was the common one in 11 Trichoderma species,other bands were polymorphic,93.5 %. Futhermore, the effect of nutrition conditions, fungicides, soil factors etc. On Trichoderma was greatly different. 2 obtained strains which were strong adaptation, would be good bio-control strains.

  16. Involvement of nitrate reductase and pyoverdine in competitiveness of Pseudomonas fluorescens strain C7R12 in soil.

    Science.gov (United States)

    Mirleau, P; Philippot, L; Corberand, T; Lemanceau, P

    2001-06-01

    Involvement of nitrate reductase and pyoverdine in the competitiveness of the biocontrol strain Pseudomonas fluorescens C7R12 was determined, under gnotobiotic conditions, in two soil compartments (bulk and rhizosphere soil), with the soil being kept at two different values of matric potential (-1 and -10 kPa). Three mutants affected in the synthesis of either the nitrate reductase (Nar(-)), the pyoverdine (Pvd(-)), or both (Nar(-) Pvd(-)) were used. The Nar(-) and Nar(-) Pvd(-) mutants were obtained by site-directed mutagenesis of the wild-type strain and of the Pvd(-) mutant, respectively. The selective advantage given by nitrate reductase and pyoverdine to the wild-type strain was assessed by measuring the dynamic of each mutant-to-total-inoculant (wild-type strain plus mutant) ratio. All three mutants showed a lower competitiveness than the wild-type strain, indicating that both nitrate reductase and pyoverdine are involved in the fitness of P. fluorescens C7R12. The double mutant presented the lowest competitiveness. Overall, the competitive advantages given to C7R12 by nitrate reductase and pyoverdine were similar. However, the selective advantage given by nitrate reductase was more strongly expressed under conditions of lower aeration (-1 kPa). In contrast, the selective advantage given by nitrate reductase and pyoverdine did not differ in bulk and rhizosphere soil, indicating that these bacterial traits are not specifically involved in the rhizosphere competence but rather in the saprophytic ability of C7R12 in soil environments. PMID:11375173

  17. Diffraction tomography of strain

    OpenAIRE

    Lionheart, W R B; Withers, P. J.

    2015-01-01

    We consider whether it is possible to recover the three dimensional strain field tomographically from neutron and x-ray diffraction data for polycrystalline materials. We show that the distribution of strain transverse to a ray cannot be deduced from one diffraction pattern accumulated along that path, but that a certain moment of that data corresponds to the transverse ray transform of the strain tensor and so may be recovered by inverting that transform given sufficient data. We show that t...

  18. Caracterización de películas biodegradables con la incorporación de un agente de biocontrol

    OpenAIRE

    TORREGROSA MORENO, BELÉN

    2016-01-01

    [ES] El control biológico es una de las alternativas más eficientes y prácticas a los fungicidas químicos para el control de enfermedades producidas por hongos en agricultura. Su aplicación práctica no ha tenido lugar en gran medida por la sensibilidad de los agentes de biocontrol (ABCs) a condiciones ambientales adversas, que dan lugar a variabilidad y poca reproducibilidad en los resultados. Se ha trabajado con el ABC Candida sake, una levadura efectiva en el control de enfer...

  19. Characterization of PhlG, a Hydrolase That Specifically Degrades the Antifungal Compound 2,4-Diacetylphloroglucinol in the Biocontrol Agent Pseudomonas fluorescens CHA0

    OpenAIRE

    Bottiglieri, Mélanie; Keel, Christoph

    2006-01-01

    The potent antimicrobial compound 2,4-diacetylphloroglucinol (DAPG) is a major determinant of biocontrol activity of plant-beneficial Pseudomonas fluorescens CHA0 against root diseases caused by fungal pathogens. The DAPG biosynthetic locus harbors the phlG gene, the function of which has not been elucidated thus far. The phlG gene is located upstream of the phlACBD biosynthetic operon, between the phlF and phlH genes which encode pathway-specific regulators. In this study, we assigned a func...

  20. 球毛壳ND35对植物生长的影响及其生防效果%Effect of Chaetomium globosum ND35 on Plant Growth and Preliminary Study of Its Biocontrol Efficacy

    Institute of Scientific and Technical Information of China (English)

    余新燕; 孟庆果; 任思栋; 刘振叶; 王茂云; 高克祥

    2009-01-01

    [Objective] The aim was to study effect of Chaetomium globosum ND35 on plant growth and preliminary study of its biocontrol efficacy, and provide basis for popularization and application of this strain. [Method] With endophytic fungus C.globosum ND35 as a tested strain, effect of C.globosum ND35 on plant growth and its biocontrol on five plant diseases were investigated in the greenhouse and field. [Result] The results showed that ND35 promoted growth of lateral root and diameter of breast height of poplar. ND35 can induce poplar to resist Poplar Valsa Canker caused by Valsa sordida and Poplar Rust caused by Melampsora puplicola. ND35 was also able to induce tomato and bean to resist Botrytis cinera. Biocontrol of Bean Stem Rot Rhizoctonia by ND35 was effective as well. [Conclusion] Induced systemic resistance by endophytic C.globosum ND35 plays an important role in biocontrol of plant diseases.%[目的]研究球毛壳ND35对植物生长的影响及其生防效果,为该菌的推广应用提供依据.[方法]以植物内生真菌球毛壳ND35为供试菌株,病原菌有杨树腐烂病菌、立枯丝核菌、番茄灰霉菌.(a)调查球毛壳ND35对植物生长的影响.在扦插杨树枝条时,设2个处理:①接种10 g长有球毛壳ND35的麦粒培养基;②接种10 g无球毛壳ND35的麦粒培养基为对照.出芽后,测量杨树高度、1.3 m处的胸径,同时调查侧根(d≥8 mm)数量.(b)大田试验测定球毛壳ND35对杨树腐烂病的拮抗作用.采用烫伤接种的方法,在主干距地面高50 cm和100 cm(约树干的1/3和2/3)处先用火烫伤树皮,再接种含病原菌菌丝体的PDA菌饼.在随后的1个多月统计发病情况,以29 d后发病的为0级,26~28 d发病的为1级,23~25 d发病的为2级,20~22 d发病的为3级,19 d之内发病的为4级.分级计数,计算发病率、病情指数和拮抗效果.(c)大田试验调查球毛壳ND35对杨树叶锈病的拮抗作用.在杨树1.3、1.7、2.1 m处分别调查2个处理组杨

  1. Strained Silicon Photonics

    Directory of Open Access Journals (Sweden)

    Ralf B. Wehrspohn

    2012-05-01

    Full Text Available A review of recent progress in the field of strained silicon photonics is presented. The application of strain to waveguide and photonic crystal structures can be used to alter the linear and nonlinear optical properties of these devices. Here, methods for the fabrication of strained devices are summarized and recent examples of linear and nonlinear optical devices are discussed. Furthermore, the relation between strain and the enhancement of the second order nonlinear susceptibility is investigated, which may enable the construction of optically active photonic devices made of silicon.

  2. Strain-engineered MOSFETs

    CERN Document Server

    Maiti, CK

    2012-01-01

    Currently strain engineering is the main technique used to enhance the performance of advanced silicon-based metal-oxide-semiconductor field-effect transistors (MOSFETs). Written from an engineering application standpoint, Strain-Engineered MOSFETs introduces promising strain techniques to fabricate strain-engineered MOSFETs and to methods to assess the applications of these techniques. The book provides the background and physical insight needed to understand new and future developments in the modeling and design of n- and p-MOSFETs at nanoscale. This book focuses on recent developments in st

  3. Influence of container design on predation rate of potential biocontrol agent, Toxorhynchites splendens (Diptera: Culicidae) against dengue vector.

    Science.gov (United States)

    Mohamad, N; Zuharah, W F

    2014-03-01

    Toxorhynchites splendens larvae are a natural predator of dengue vector mosquito larvae, Aedes albopictus. This study was carried out to evaluate the predation rate of Tx. splendens third instar larvae on Ae. albopictus larvae in 24 h. Each predator was offered prey at a density between 10 to 50 individuals. Predation rate of Tx. splendens were also tested with two manipulated factors; various types of container and different water volumes. The experiment was evaluated in man-made containers (tin cans, plastic drinking glasses and rubber tires) and natural container (bamboo stumps) which were filled with different water volumes (full, half full, 1/4 full, and 1/8 full). The prey density and the characteristics of the container were found as significant factors which influence the predation rate of Tx. splendens. The predator consumed significantly more prey at higher prey densities (40 and 50 preys) compared to the lowest density (10 preys) (F=3.935, df=4, p=0.008). The results showed significantly higher consumption in horizontal shaped container of rubber tire than in vertical shape of bamboo stumps (F=3.100, df=3, p=0.029). However, the water volume had no significant effect on predation rate of Tx. splendens (F=1.736, df=3, p=0.162). We generally suggest that Tx. splendens is best to be released in discarded tires or any other containers with horizontal shape design with wide opening since Tx. splendens can become more effective in searching prey in this type of container design. This predator is also a suitable biocontrol candidates to be introduced either in wet and dry seasons in Malaysia.

  4. Do biopesticides affect the demographic traits of a parasitoid wasp and its biocontrol services through sublethal effects?

    Directory of Open Access Journals (Sweden)

    Antonio Biondi

    Full Text Available Pesticide risk assessments are usually based on short-term acute toxicity tests, while longer-term population dynamic related traits, critical to the success of biological control and Integrated Pest Management (IPM programs, are often overlooked. This is increasingly important with respect to new biopesticides that frequently cause no short-term acute effects, but that can induce multiple physiological and behavioral sublethal effects, leading to a decrease in population growth and ecosystem services. In this study we assessed the lethal and sublethal effects of six biopesticides [abamectin, azadirachtin, Bacillus thuringiensis, borax plus citrus oil (Prev-Am®, emamectin benzoate, and spinosad], used in tomato crops to control the invasive pest Tuta absoluta (Lepidoptera: Gelechiidae, on adults and pupae of the parasitoid Bracon nigricans (Hymenoptera: Braconidae. Data on female survival and production of female offspring were used to calculate population growth indexes as a measure of population recovery after pesticide exposure. Spinosad caused 100% and 80% mortality in exposed adults (even 10 d after the treatment and pupae, respectively. Although most of the biopesticides had low levels of acute toxicity, multiple sublethal effects were observed. The biocontrol activity of both females that survived 1-h and 10-d old residues, and females that emerged from topically treated pupae was significantly affected by the application of the neurotoxic insecticides emamectin benzoate and abamectin. Furthermore, very low B. nigricans demographic growth indices were estimated for these two insecticides, indicating potential local extinction of the wasp populations. Among the tested products, Bt proved to be the safest for B. nigricans adults and pupae. Our findings emphasize that acute toxicity assessment alone cannot fully predict the actual impact of pesticides on non-target parasitoids. Thus, sublethal effects related to the species specific life

  5. Biosynthesis of Antibiotic Leucinostatins in Bio-control Fungus Purpureocillium lilacinum and Their Inhibition on Phytophthora Revealed by Genome Mining

    Science.gov (United States)

    Li, Erfeng; Mao, Zhenchuan; Ling, Jian; Yang, Yuhong; Yin, Wen-Bing; Xie, Bingyan

    2016-01-01

    Purpureocillium lilacinum of Ophiocordycipitaceae is one of the most promising and commercialized agents for controlling plant parasitic nematodes, as well as other insects and plant pathogens. However, how the fungus functions at the molecular level remains unknown. Here, we sequenced two isolates (PLBJ-1 and PLFJ-1) of P. lilacinum from different places Beijing and Fujian. Genomic analysis showed high synteny of the two isolates, and the phylogenetic analysis indicated they were most related to the insect pathogen Tolypocladium inflatum. A comparison with other species revealed that this fungus was enriched in carbohydrate-active enzymes (CAZymes), proteases and pathogenesis related genes. Whole genome search revealed a rich repertoire of secondary metabolites (SMs) encoding genes. The non-ribosomal peptide synthetase LcsA, which is comprised of ten C-A-PCP modules, was identified as the core biosynthetic gene of lipopeptide leucinostatins, which was specific to P. lilacinum and T. ophioglossoides, as confirmed by phylogenetic analysis. Furthermore, gene expression level was analyzed when PLBJ-1 was grown in leucinostatin-inducing and non-inducing medium, and 20 genes involved in the biosynthesis of leucionostatins were identified. Disruption mutants allowed us to propose a putative biosynthetic pathway of leucinostatin A. Moreover, overexpression of the transcription factor lcsF increased the production (1.5-fold) of leucinostatins A and B compared to wild type. Bioassays explored a new bioactivity of leucinostatins and P. lilacinum: inhibiting the growth of Phytophthora infestans and P. capsici. These results contribute to our understanding of the biosynthetic mechanism of leucinostatins and may allow us to utilize P. lilacinum better as bio-control agent. PMID:27416025

  6. Can the halophilic ciliate Fabrea salina be used as a bio-control of microalgae blooms in solar salterns?

    Science.gov (United States)

    Hong, Hyun Pyo; Choi, Joong Ki

    2015-09-01

    The microlage Dunaliella salina, a major producer in salterns, is a serious problem for salt production. In this study we tried to assess if Fabrea salina can control D. salina. By parameterising numerical and functional response (growth and grazing vs prey abundance, respectively) at 90 psu and 30°C, where the ciliate is abundant and grows well, we developed a predator-prey model. The model is used to explore how change in microalga growth rate affect the dynamics, and the functional response is used in combination with field data to assess the potential impact of F. salina on D. salina. Over the 20 d simulation the ciliate controlled the prey population under all prey growth rates; although once D. salina were exhausted below the threshold level, F. salina died due to starvation, allowing the alga to increase in abundance, resulting in one or two predatorprey cycle, depending on prey growth rate. In general, the model predicted trends observed by others in the field, suggesting that it provided a good prediction of what may occur under the conditions we examined. Likewise we show that the ciliate can have a high impact on microalgal populations in the field. Finally, a literature review indicated that F. salina could be a good competitor with other protozoa and metazoan in salterns, depending on salinity and temperature, which requires further study and attention. In summary, we encourage continued studies on this unique ciliate on solar salterns and suggest that it may be useful in the bio-control of micoalgae.

  7. Biosynthesis of Antibiotic Leucinostatins in Bio-control Fungus Purpureocillium lilacinum and Their Inhibition on Phytophthora Revealed by Genome Mining.

    Directory of Open Access Journals (Sweden)

    Gang Wang

    2016-07-01

    Full Text Available Purpureocillium lilacinum of Ophiocordycipitaceae is one of the most promising and commercialized agents for controlling plant parasitic nematodes, as well as other insects and plant pathogens. However, how the fungus functions at the molecular level remains unknown. Here, we sequenced two isolates (PLBJ-1 and PLFJ-1 of P. lilacinum from different places Beijing and Fujian. Genomic analysis showed high synteny of the two isolates, and the phylogenetic analysis indicated they were most related to the insect pathogen Tolypocladium inflatum. A comparison with other species revealed that this fungus was enriched in carbohydrate-active enzymes (CAZymes, proteases and pathogenesis related genes. Whole genome search revealed a rich repertoire of secondary metabolites (SMs encoding genes. The non-ribosomal peptide synthetase LcsA, which is comprised of ten C-A-PCP modules, was identified as the core biosynthetic gene of lipopeptide leucinostatins, which was specific to P. lilacinum and T. ophioglossoides, as confirmed by phylogenetic analysis. Furthermore, gene expression level was analyzed when PLBJ-1 was grown in leucinostatin-inducing and non-inducing medium, and 20 genes involved in the biosynthesis of leucionostatins were identified. Disruption mutants allowed us to propose a putative biosynthetic pathway of leucinostatin A. Moreover, overexpression of the transcription factor lcsF increased the production (1.5-fold of leucinostatins A and B compared to wild type. Bioassays explored a new bioactivity of leucinostatins and P. lilacinum: inhibiting the growth of Phytophthora infestans and P. capsici. These results contribute to our understanding of the biosynthetic mechanism of leucinostatins and may allow us to utilize P. lilacinum better as bio-control agent.

  8. Biocontrol and Plant Growth Promotion Characterization of Bacillus Species Isolated from Calendula officinalis Rhizosphere.

    Science.gov (United States)

    Ait Kaki, Asma; Kacem Chaouche, Noreddine; Dehimat, Laid; Milet, Asma; Youcef-Ali, Mounia; Ongena, Marc; Thonart, Philippe

    2013-12-01

    The phenotypic and genotypic diversity of the plant growth promoting Bacillus genus have been widely investigated in the rhizosphere of various agricultural crops. However, to our knowledge this is the first report on the Bacillus species isolated from the rhizosphere of Calendula officinalis. 15 % of the isolated bacteria were screened for their important antifungal activity against Fusarium oxysporum, Botrytis cinerea, Aspergillus niger, Cladosporium cucumerinium and Alternaria alternata. The bacteria identification based on 16S r-RNA and gyrase-A genes analysis, revealed strains closely related to Bacillus amyloliquefaciens, B. velezensis, B. subtilis sub sp spizezenii and Paenibacillus polymyxa species. The electro-spray mass spectrometry coupled to liquid chromatography (ESI-LC MS) analysis showed that most of the Bacillus isolates produced the three lipopeptides families. However, the P. polymyxa (18SRTS) didn't produce any type of lipopeptides. All the tested Bacillus isolates produced cellulase but the protease activity was observed only in the B. amyloliquefaciens species (9SRTS). The Salkowsky colorimetric test showed that the screened bacteria synthesized 6-52 μg/ml of indole 3 acetic acid. These bacteria produced siderophores with more than 10 mm wide orange zones on chromazurol S. The greenhouse experiment using a naturally infested soil with Sclerotonia sclerotiorum showed that the B. amyloliquefaciens (9SRTS) had no significant (P > 0.05) effect on the pre-germination of the chickpea seeds. However, it increased the size of the chickpea plants and reduced the stem rot disease (P officinalis and other crop systems. PMID:24426149

  9. Bioprotection of Spruce Logs Against Sapstain Using an Albino Strain of Ceratocystis resinifera.

    Science.gov (United States)

    Morin, Chantal; Tanguay, Philippe; Breuil, Colette; Yang, Dian-Qing; Bernier, Louis

    2006-05-01

    ABSTRACT We recovered a spontaneous albino strain from ascospores of Ceratocystis resinifera, a sapstain fungus that grows deeply and rapidly in freshly felled conifer trees. This albino strain, named Kasper, was tested for its ability to prevent discoloration of spruce sapwood caused by wild-type sapstain fungi and compared with Cartapip 97, a commercially available biological control agent of sapstain in lodgepole pine and red pine logs. In a laboratory trial, Kasper reduced sapstain of white spruce logs as much as 94.4% and was more efficient than Cartapip 97. In field trials conducted in an area north of Québec City, Kasper reduced sapstain of black spruce as much as 80%. In three of four field trials, Kasper was significantly more efficient than Cartapip 97 in reducing sapstain development. The exception was encountered in a 2003 trial conducted in a sawmill yard where Kasper did not reduce sapstain. In a field trial conducted in western Canada, at Aleza Lake forest near Prince George, Kasper almost totally prevented the development of sapstain, even after 24 weeks. These results suggest albino strains derived from C. resinifera might be an additional source of potential biocontrol agents against sapstain.

  10. Histopathological studies of sclerotia of phytopathogenic fungi parasitized by a GFP transformed Trichoderma virens antagonistic strain.

    Science.gov (United States)

    Sarrocco, Sabrina; Mikkelsen, Lisbeth; Vergara, Mariarosaria; Jensen, Dan Funck; Lübeck, Mette; Vannacci, Giovanni

    2006-02-01

    The gfp gene from the jellyfish Aequorea victoria, coding for the Green Fluorescent Protein (GFP), was used as a reporter gene to transform a Trichoderma virens strain I10, characterized as having a promising biocontrol activity against a large number of phytopathogenic fungi. On the basis of molecular and biological results, a stable GFP transformant was selected for further experiments. In order to evaluate the effects of GFP transformation on mycoparasitic ability of T. virens I10, sclerotia of Sclerotium rolfsii, Sclerotinia sclerotiorum and S. minor were inoculated with the T. virens strain I10 GFP transformant or the wild type strain. Statistical analysis of percentages of decayed sclerotia showed that the transformation of the antagonistic isolate with the GFP reporter gene did not modify mycoparasitic activity against sclerotia. Sclerotium colonization was followed by fluorescent microscopy revealing intracellular growth of the antagonist in the cortex (S. rolfsii) and inter-cellular growth in the medulla (S. rolfsii, and S. sclerotiorum). The uniformly distributed mycelium of T. virens just beneath the rind of sclerotia of both S. rolfsii and S. sclerotiorum suggests that the sclerotia became infected at numerous randomly distributed locations without any preferential point of entry. PMID:16388938

  11. Biological control of grape crown gall by strain f2/5 is not associated with agrocin production or competition for attachment sites on grape cells.

    Science.gov (United States)

    Burr, T J; Reid, C L; Tagliati, E; Bazzi, C; Süle, S

    1997-07-01

    ABSTRACT Agrocin-minus mutants of nontumorigenic Agrobacterium vitis strain F2/5 controlled grape crown gall as well as the wild-type strain, indicating that agrocin is not a major factor in the mechanism of biological control. Relative levels of attachment to grape cells by tumorigenic and biocontrol strains were also measured. Attachment of tumorigenic strains (CG49 and K306) and biological control strains (F2/5 and agrocin-minus mutant 1077) was often reduced when mixtures of the strains were applied. However, high populations (10(3) to 10(5) CFU/ml) of all strains attached following mixed inoculations, suggesting that competition for attachment sites is also not a factor in the mechanism of biological control. Transfer of T-DNA to grape by CG49 was prevented or greatly inhibited in the presence of F2/5 or 1077 as measured by expression of the GUS reporter gene. The Ti plasmid virulence genes, however, were induced by exudates from grape shoots that had been inoculated with F2/5. Sonicated and autoclaved preparations of F2/5 and 1077 did not control crown gall or inhibit T-DNA transfer. Control by F2/5 is specific to grape, since gall formation on tomato, sunflower, and Kalanchoe daigremontiana were not inhibited. PMID:18945092

  12. Isolation and characterization of a new Bacillus thuringiensis strain with a promising toxicity against Lepidopteran pests.

    Science.gov (United States)

    Boukedi, Hanen; Sellami, Sameh; Ktari, Sonia; Belguith-Ben Hassan, Najeh; Sellami-Boudawara, Tahya; Tounsi, Slim; Abdelkefi-Mesrati, Lobna

    2016-01-01

    Insecticides derived from Bacillus thuringiensis are gaining worldwide importance as environmentally desirable alternatives to chemicals for the control of pests in public health and agriculture. Isolation and characterization of new strains with higher and broader spectrum of activity is an ever growing field. In the present work, a novel Tunisian B. thuringiensis isolate named BLB459 was characterized and electrophoresis assay showed that among a collection of 200 B. thuringiensis strains, the plasmid profile of BLB459 was distinctive. SmaI-PFGE typing confirmed the uniqueness of the DNA pattern of this strain, compared with BUPM95 and HD1 reference strains. PCR and sequencing assays revealed that BLB459 harbored three cry genes (cry30, cry40 and cry54) corresponding to the obtained molecular sizes in the protein pattern. Interestingly, PCR-RFLP assay demonstrated the originality of the BLB459 cry30-type gene compared to the other published cry30 genes. Insecticidal bioassays showed that BLB459 spore-crystal suspension was highly toxic to both Ephestia kuehniella and Spodoptera littoralis with LC50 values of about 64 (53-75) and 80 (69-91) μg of toxin cm(-2), respectively, comparing with that of the commercial strain HD1 used as reference. Important histopathological effects of BLB459 δ-endotoxins on the two tested larvae midguts were detected, traduced by the vacuolization of the apical cells, the damage of microvilli, and the disruption of epithelial cells. These results proved that BLB459 strain could be of a great interest for lepidopteran biocontrol. PMID:27242138

  13. Development of a powder formulation based on Bacillus cereus sensu lato strain B25 spores for biological control of Fusarium verticillioides in maize plants.

    Science.gov (United States)

    Martínez-Álvarez, Juan C; Castro-Martínez, Claudia; Sánchez-Peña, Pedro; Gutiérrez-Dorado, Roberto; Maldonado-Mendoza, Ignacio E

    2016-05-01

    Maize is an economically important crop in northern Mexico. Different fungi cause ear and root rot in maize, including Fusarium verticillioides (Sacc.) Nirenberg. Crop management of this pathogen with chemical fungicides has been difficult. By contrast, the recent use of novel biocontrol strategies, such as seed bacterization with Bacillus cereus sensu lato strain B25, has been effective in field trials. These approaches are not without their problems, since insufficient formulation technology, between other factors, can limit success of biocontrol agents. In response to these drawbacks, we have developed a powder formulation based on Bacillus B25 spores and evaluated some of its characteristics, including shelf life and efficacy against F. verticillioides, in vitro and in maize plants. A talc-based powder formulation containing 1 × 10(9) c.f.u. g(-1) was obtained and evaluated for seed adherence ability, seed germination effect, shelf life and antagonism against F. verticillioides in in vitro and in planta assays. Seed adherence of viable bacterial spores ranged from 1.0 to 1.41 × 10(7) c.f.u. g(-1). Bacteria did not display negative effects on seed germination. Spore viability for the powder formulation slowly decreased over time, and was 53 % after 360 days of storage at room temperature. This formulation was capable of controlling F. verticillioides in greenhouse assays, as well as eight other maize phytopathogenic fungi in vitro. The results suggest that a talc-based powder formulation of Bacillus B25 spores may be sufficient to produce inoculum for biocontrol of maize ear and root rots caused by F. verticillioides. PMID:27038945

  14. Genomics of Three New Bacteriophages Useful in the Biocontrol of Salmonella.

    Science.gov (United States)

    Bardina, Carlota; Colom, Joan; Spricigo, Denis A; Otero, Jennifer; Sánchez-Osuna, Miquel; Cortés, Pilar; Llagostera, Montserrat

    2016-01-01

    Non-typhoid Salmonella is the principal pathogen related to food-borne diseases throughout the world. Widespread antibiotic resistance has adversely affected human health and has encouraged the search for alternative antimicrobial agents. The advances in bacteriophage therapy highlight their use in controlling a broad spectrum of food-borne pathogens. One requirement for the use of bacteriophages as antibacterials is the characterization of their genomes. In this work, complete genome sequencing and molecular analyses were carried out for three new virulent Salmonella-specific bacteriophages (UAB_Phi20, UAB_Phi78, and UAB_Phi87) able to infect a broad range of Salmonella strains. Sequence analysis of the genomes of UAB_Phi20, UAB_Phi78, and UAB_Phi87 bacteriophages did not evidence the presence of known virulence-associated and antibiotic resistance genes, and potential immunoreactive food allergens. The UAB_Phi20 genome comprised 41,809 base pairs with 80 open reading frames (ORFs); 24 of them with assigned function. Genome sequence showed a high homology of UAB_Phi20 with Salmonella bacteriophage P22 and other P22likeviruses genus of the Podoviridae family, including ST64T and ST104. The DNA of UAB_Phi78 contained 44,110 bp including direct terminal repeats (DTR) of 179 bp and 58 putative ORFs were predicted and 20 were assigned function. This bacteriophage was assigned to the SP6likeviruses genus of the Podoviridae family based on its high similarity not only with SP6 but also with the K1-5, K1E, and K1F bacteriophages, all of which infect Escherichia coli. The UAB_Phi87 genome sequence consisted of 87,669 bp with terminal direct repeats of 608 bp; although 148 ORFs were identified, putative functions could be assigned to only 29 of them. Sequence comparisons revealed the mosaic structure of UAB_Phi87 and its high similarity with bacteriophages Felix O1 and wV8 of E. coli with respect to genetic content and functional organization. Phylogenetic analysis of large

  15. Genomics of three new bacteriophages useful in the biocontrol of Salmonella

    Directory of Open Access Journals (Sweden)

    Carlota eBardina

    2016-04-01

    Full Text Available Non-typhoid Salmonella is the principal pathogen related to food-borne diseases throughout the world. Widespread antibiotic resistance has adversely affected human health and has encouraged the search for alternative antimicrobial agents. The advances in bacteriophage therapy highlight their use in controlling a broad spectrum of food-borne pathogens. One requirement for the use of bacteriophages as antibacterials is the characterization of their genomes. In this work, complete genome sequencing and molecular analyses were carried out for three new virulent Salmonella-specific bacteriophages (UAB_Phi20, UAB_Phi78, and UAB_Phi87 able to infect a broad range of Salmonella strains. Sequence analysis of the genomes of UAB_Phi20, UAB_Phi78, and UAB_Phi87 bacteriophages did not evidence the presence of known virulence-associated and antibiotic resistance genes, and potential immunoreactive food allergens. The UAB_Phi20 genome comprised 41,809 base pairs with 80 open reading frames (ORFs; 24 of them with assigned function. Genome sequence showed a high homology of UAB_Phi20 with Salmonella bacteriophage P22 and other P22likeviruses genus of the Podoviridae family, including ST64T and ST104. The DNA of UAB_Phi78 contained 44,110 bp including direct terminal repeats of 179 bp and 58 putative ORFs were predicted and 20 were assigned function. This bacteriophage was assigned to the SP6likeviruses genus of the Podoviridae family based on its high similarity not only with SP6 but also with the K1-5, K1E, and K1F bacteriophages, all of which infect Escherichia coli. The UAB_Phi87 genome sequence consisted of 87,669 bp with terminal direct repeats of 608 bp; although 148 ORFs were identified, putative functions could be assigned to only 29 of them. Sequence comparisons revealed the mosaic structure of UAB_Phi87 and its high similarity with bacteriophages Felix O1 and wV8 of E. coli with respect to genetic content and functional organization. Phylogenetic

  16. Proteins Related to the Biocontrol of Pythium Damping-off in Maize with Trichoderma harzianum Rifai

    Institute of Scientific and Technical Information of China (English)

    Jie CHEN; Gary E HARMAN; Afio COMIS; Gen-Wu CHENG

    2005-01-01

    Induced resistance has been evidenced as one of mechanisms of Trichoderma to control plantdiseases, however, no study showed the change of host proteomics in Trichoderma-induced resistance ofmaize against damping-off caused by Pythium ultimum Trow. The mechanism of Trichoderma harzianumRifai for controlling maize seedling disease caused by Pythium ultimum Trow was investigated firstly byproteome technique and the result suggested that T. harzianum strain T22 was not only able to promoteseedling growth but also protein accumulation. One-dimensional electrophoresis assay showed that morebands appeared on the gel with T22 or T22 combined with P. ultimum (T22 + P. ultimum) treatment than withother treatments. Enzyme assay showed that two chitinases of the root sample were more activated in thetreatments with T22 than in the other treatments without T22. Proteins in the seedling roots from the varioustreatments were separated through protein extraction and 2-D electrophoresis technique. In the seedlingsproduced from the T22-treated seeds, there were 104 up-regulated proteins and 164 down-regulated pro-teins relative to the control, and 97 and 150, respectively, after treatment with T22 + P. ultimum; however,with P. ultimum alone the values were much lower than with the other two treatments. The correlationcoefficient values were 0.72, 0.51 and 0.49 for the comparison of protein spot distribution on gel amongcontrol with T22, P. ultimum and T22 + P. ultimum, respectively. So it seemed that P. ultimum infection wasmore effective than T22 in interfering with the host proteome profile. Furthermore, analysis with MALDI-TOF-MAS showed that some important proteins associated with defensive reactions were identified in T22or T22 + P. ultimum treatments, including endochitinase, pathogenesis-related protein PRMS (pathogenesis-related maize seed), GTP-binding protein, isoflavone reductase and other proteins related to respiration. Allthose proteins are probably part of the

  17. Proteomics related to the biocontrol of Pythium damping off in maize with Trichoderma harzianum

    Institute of Scientific and Technical Information of China (English)

    CHEN Jie; Gary G Harman; Alfio Comis

    2004-01-01

    @@ Trichoderma harzianum strain T22 controls various diseases of maize and other crops, including seedling and root rots caused by Pythium ultimum. Seedlings of inbred line Mo17 were grown from T22-treated or untreated seeds in field soil or in field soil intested with the pathogen. Five days after planting, seedlings of Mo17 (5-days-old) were smaller in the presence of P. ultimum and larger in the presence of T22 relative to the control. The combination of T22 with P. ultimum (T22 + P.ultimum ) resulted in plants as large as T22 alone. Methods for protein extraction and 2-D gel electrophoresis were developed. Proteins in seedlings roots from the various treatments were separated on 2-D gels and analyzed using PDQuestTM 2-D software. With seedlings produced from T22-treated seeds, there were 104 unmatched proteins and 164 matched proteins relative to the control, and 97 and 150 from the treatment with T22 + P. ultimum, respectively, however, with P. ultimum alone the numbers were much lower than above two treatments. Comparatively, there was very lower similarity of proteome patterns of seedling roots with T22 or P. ultimum or both to control seedlings,the correlative coefficient values were 0.72, 0.51 and 0.49 for the comparisons among control with T22, P. ultimum and T22 + P. ultimum, respectively. Moreover, correlative coefficient of proteome patterns between T22 with P. ultimum was only 0.65, and T22 fungal proteome were also not same as any one of seedling roots with various treatments. Taken together, the components in seedling root proteome seemed to be mostly coming from Mo17 plants themselves and affected strongly by either microbes, but the effects appeared to be stronger by P. ultimum than by T22. 41 spots were selected for protein mass fingerprinting identification, and most detected-spots were intensified in abundance by T22 or T22 + P. ultimum treatments such as pathogenesis-related protein and endochitnase etc. SOD (Mn) was found to be involved in

  18. Highly Strained Organophosphorus Compounds

    NARCIS (Netherlands)

    Slootweg, J.C.

    2005-01-01

    In our research on small, strained organophosphorus ring systems we became interested in the synthesis and applications of species that are even more strained than the parent phosphirane, by introducing an exocyclic double bond (methylenephosphirane), and by cyclopropyl spirofusion to the edge (e.g

  19. Variations on conserved signaling pathways in biocontrol and development:G protein and MAPK genes of Trichoderma.atroviride and T.virens

    Institute of Scientific and Technical Information of China (English)

    Benjiamin A Horwitz

    2004-01-01

    @@ Filamentous fungi employ conserved eukaryotic signaling pathway to detect and respond to environmental signals, including the presence of the host. Genetic experiment in which a particular signaling protein is lost, or its activity enhanced, have defined some of the function of heterotrimeric G proteins and MAP kinases in development and virulence. A hallmark of these studies is that orthologs in different species may have different functions. Antagonistic fungal-fungal interactions form the basis for biological control of plant disease. These interactions may employ novel modes of regulation by conserved signaling elements. Tag1, a G protein a subunit of Trichoderma. atroviride belonging to fungal Gi class, is involved in repression of sporulation and hyphal coiling(1). Deletion of ortholog of this gene, TgaA, in Trichoderma (Gliocladium) virens, however, did not affect sporulation and growth, yet tgaA mutants are unable to parasitize S. rolfsii sclerotia(2). Mutation of a second G αsubunit gene is now under study. TmkA, a MAPK gene of T. virens, is involved in biocontrol properties and repression of conidiation (3). Using suppression-subtraction hybridization and other approaches, we are beginning to identify additional elements of the signaling cascades and their downsteam targets. The role of G protein and MAPK genes are sometimes specific to a particular host fungus or to parasitism of mycelia or sclerotia (2, 3). Also of relevance to biocontrol, signal transduction pathway provide a means to alter the balance between sporulation, mycelial growth and hyphal coiling.

  20. Biocontrol of Potato Common Scab is Associated with High Pseudomonas fluorescens LBUM223 Populations and Phenazine-1-Carboxylic Acid Biosynthetic Transcript Accumulation in the Potato Geocaulosphere.

    Science.gov (United States)

    Arseneault, Tanya; Goyer, Claudia; Filion, Martin

    2016-09-01

    Pseudomonads are often used as biocontrol agents because they display a broad range of mechanisms to control diseases. Common scab of potato, caused by Streptomyces scabies, was previously reported to be controlled by Pseudomonas fluorescens LBUM223 through phenazine-1-carboxylic acid (PCA) production. In this study, we aimed at characterizing the population dynamics of LBUM223 and the expression of phzC, a key gene involved in the biosynthesis of PCA, in the rhizosphere and geocaulosphere of potato plants grown under controlled and field conditions. Results obtained from controlled experiments showed that soil populations of LBUM223 significantly declined over a 15-week period. However, at week 15, the presence of S. scabies in the geocaulosphere was associated with significantly higher populations of LBUM223 than when the pathogen was absent. It also led to the detection of significantly higher phzC gene transcript numbers. Under field conditions, soil populations of LBUM223 followed a similar decline in time when a single inoculation was applied in spring but remained stable when reinoculated biweekly, which also led to greater phzC gene transcripts accumulation. Taken together, our findings suggest that LBUM223 must colonize the potato geocaulosphere at high levels (10(7) bacteria/g of soil) in order to achieve biocontrol of common scab through increased PCA production. PMID:27088392

  1. Isolation and characterization of rhizosphere bacteria for the biocontrol of the damping-off disease of tomatoes in Tunisia.

    Science.gov (United States)

    Hammami, Inés; Ben Hsouna, Anis; Hamdi, Naceur; Gdoura, Radhouane; Triki, Mohamed Ali

    2013-01-01

    Fluorescent Pseudomonas spp., isolated from tomato and pepper plants rhizosphere soil, was evaluated in vitro as a potential antagonist of fungal pathogens. Pseudomonas strains were tested against the causal agents of tomatoes damping-off (Sclerotinia sclerotiorum), root rot (Fusarium solani), and causal agents of stem canker and leaf blight (Alternaria alternata). For this purpose, dual culture antagonism assays were carried out on 25% tryptic soy agar, King B medium and potato dextrose agar to determine the effect of the strains on mycelial growth of the pathogens. In addition, strains were screened for their ability to produce exoenzymes and siderophores. All the strains significantly inhibited Alternaria alternata, particularly in 25% TSA medium. Antagonistic effect on Sclerotinia sclerotiorum and Fusarium solani was greater on King B medium. Protease was produced by 30% of the strains, but no strain produced cellulase or chitinase. Finally, the selected Pseudomonas strain, Psf5, was evaluated on tomato seedling development and as a potential candidate for controlling tomato damping-off caused by Sclerotinia sclerotiorum, under growth chamber conditions. In vivo studies resulted in significant increases in plant stand as well as in root dry weight. Psf5 was able to establish and survive in tomato plants rhizosphere after 40days following the planting of bacterized seeds. PMID:24296079

  2. Trichodiene production in a Trichoderma harzianum erg1-silenced strain provides evidence of the importance of the sterol biosynthetic pathway in inducing plant defense-related gene expression

    Science.gov (United States)

    Trichoderma species are often used as biocontrol agents against plant-pathogenic fungi. A complex molecular interaction occurs among the biocontrol agent, the antagonistic fungus, and the plant. Terpenes and sterols produced by the biocontrol fungus have been found to affect gene expression in both ...

  3. Characterization of endophytic Bacillus strains from tomato plants (Lycopersicon esculentum) displaying antifungal activity against Botrytis cinerea Pers.

    Science.gov (United States)

    Kefi, Asma; Ben Slimene, Imen; Karkouch, Ines; Rihouey, Christophe; Azaeiz, Sana; Bejaoui, Marwa; Belaid, Rania; Cosette, Pascal; Jouenne, Thierry; Limam, Ferid

    2015-12-01

    Eighty endophytic bacteria were isolated from healthy tissues of roots, stems, leaves and fruits of tomato plants (Lycopersicon esculentum). Four strains, named BL1, BT5, BR8 and BF11 were selected for their antagonism against Botrytis cinerea, a phytopathogenic fungus responsible of gray mold in several important crops, with growth inhibitory activity ranging from 27 to 53%. Morphological, biochemical, and molecular parameters as 16S rDNA sequencing demonstrated that the selected bacterial strains were related to Bacillus species which are known to produce and secrete a lot of lipopeptides with strong inhibitory effect against pathogen mycelial growth. Electrospray mass spectrometry analysis showed that these strains produced heterogeneous mixture of antibiotics belonging to fengycin and surfactin for BL1 and BT5, to iturin and surfactin for BR8, to bacillomycin D, fengycin and surfactin for BF11. Furthermore, these bacteria exhibited biocontrol potential by reducing the disease severity when tested on detached leaflets. Based on their antifungal activity against Botrytis cinerea, these strains could be used for biological control of plant diseases.

  4. In vitro and in vivo antagonism of pathogenic turfgrass fungi by Streptomyces hygroscopicus strains YCED9 and WYE53.

    Science.gov (United States)

    Chamberlain, K; Crawford, D L

    1999-07-01

    Disease prevention is a current practice used to minimize fungal diseases of turfgrasses in lawns and golf greens. Prevention is accomplished through fungicide applications, and by periodic thatch removal. During the development of a microbial biodethatch product utilizing the lignocellulose-degrading Streptomyces hygroscopicus strains YCED9 and WYE53, we demonstrated using in vitro plate antagonism bioassays that both strains are antagonists of various turfgrass fungal pathogens. These activities were present when the cultures were growing on thatch, as demonstrated by antifungal antagonism bioassays with culture filtrates. Experiments conducted using a growth chamber demonstrated that a bio-dethatch formulation containing spores of strains YCED9 and WYE53 in a zeolite carrier, provided protection for Kentucky bluegrass seedlings against turfgrass pathogens, including Pythium ultimum, Fusarium oxysporum, Rhizoctonia solani, Sclerotinia homeocarpa, Gaeumannomyces graminis and Microdochium nivale. Results showed that by integrating the use of the S. hygroscopicus YCED9/WYE53 bio-dethatch formulation into routine turf management practices, it should be possible to both minimize thatch build-up while also controlling fungal turfgrass diseases by way of the antifungal biocontrol activity of these strains. This in turn would help control fungal pathogens in turfgrass while minimizing the need for routine chemical fungicide applications. PMID:10455494

  5. Characterization of endophytic Bacillus strains from tomato plants (Lycopersicon esculentum) displaying antifungal activity against Botrytis cinerea Pers.

    Science.gov (United States)

    Kefi, Asma; Ben Slimene, Imen; Karkouch, Ines; Rihouey, Christophe; Azaeiz, Sana; Bejaoui, Marwa; Belaid, Rania; Cosette, Pascal; Jouenne, Thierry; Limam, Ferid

    2015-12-01

    Eighty endophytic bacteria were isolated from healthy tissues of roots, stems, leaves and fruits of tomato plants (Lycopersicon esculentum). Four strains, named BL1, BT5, BR8 and BF11 were selected for their antagonism against Botrytis cinerea, a phytopathogenic fungus responsible of gray mold in several important crops, with growth inhibitory activity ranging from 27 to 53%. Morphological, biochemical, and molecular parameters as 16S rDNA sequencing demonstrated that the selected bacterial strains were related to Bacillus species which are known to produce and secrete a lot of lipopeptides with strong inhibitory effect against pathogen mycelial growth. Electrospray mass spectrometry analysis showed that these strains produced heterogeneous mixture of antibiotics belonging to fengycin and surfactin for BL1 and BT5, to iturin and surfactin for BR8, to bacillomycin D, fengycin and surfactin for BF11. Furthermore, these bacteria exhibited biocontrol potential by reducing the disease severity when tested on detached leaflets. Based on their antifungal activity against Botrytis cinerea, these strains could be used for biological control of plant diseases. PMID:26347324

  6. Characteristics and antimicrobial activity of Bacillus subtilis strains isolated from soil.

    Science.gov (United States)

    Todorova, Sevdalina; Kozhuharova, Lubka

    2010-07-01

    Antagonistic Bacillus strains were isolated from soil and analyzed for the purpose of determining whether they could be used as natural biological agents. Primary in vitro screening for antagonism of the isolates was performed against five phytopathogenic mould fungi. Strains TS 01 and ZR 02 exhibited the most pronounced inhibitory effects. They were identified as Bacillus subtilis on the basis of their morphological, cultural and physiology-biochemical properties as well as their hierarchical cluster analysis conducted by means of computer program SPSS. The antimicrobial activity of the strains from cultural medium and sterile filtrate were determined in vitro against a great number of predominantly phytopathogenic fungi and bacteria. TS 01 and ZR 02 strains exhibited very broad and at the same time degree varying antibiotic spectra of activities against both Gram-positive and Gram-negative microorganisms. Many of them were tested against sensitivity to the antimicrobial action of B. subtilis for the very first time. B. subtilis TS 01 and ZR 02 showed highest antifungal activity (sterile zone in diameter over 37 mm) against Alternaria solani, Botrytis cinerea, Monilia linhartiana 869, Phytophthora cryptogea 759/1 and Rhizoctonia sp. The most sensitive bacterial species were found to be Pseudomonas syringae pv. tomato Ro and Xanthomonas campestris with sterile zones 48.0 and 50.0 mm in diameter, respectively. The latter draws a conclusion that the isolated and identified Bacillus subtilis strains are promising natural biocontrol agents and should be further studied and tested for control of numerous plant diseases. PMID:24026925

  7. Sprains and Strains

    Science.gov (United States)

    ... people at risk for strains. Gymnastics, tennis, rowing, golf, and other sports that require extensive gripping can ... Trials and You was designed to help people learn more about clinical trials, why they matter, and ...

  8. The strained state cosmology

    CERN Document Server

    Tartaglia, Angelo

    2015-01-01

    Starting from some relevant facts concerning the behaviour of the universe over large scale and time span, the analogy between the geometric approach of General Relativ- ity and the classical description of an elastic strained material continuum is discussed. Extending the elastic deformation approach to four dimensions it is shown that the accelerated expansion of the universe is recovered. The strain field of space-time repro- duces properties similar to the ones ascribed to the dark energy currently called in to explain the accelerated expansion. The strain field in the primordial universe behaves as radiation, but asymptotically it reproduces the cosmological constant. Subjecting the theory to a number of cosmological tests confirms the soundness of the approach and gives an optimal value for the one parameter of the model, i.e. the bulk modulus of the space-time continuum. Finally various aspects of the Strained State Cosmology (SSC) are discussed and contrasted with some non-linear massive gravity theor...

  9. An Ultrasonic Strain Gauge

    OpenAIRE

    Kersemans, Mathias; Allaer, Klaas; Degrieck, Joris; Van Den Abeele, Koen; Pyl, Lincy; Zastavnik, Filip; Sol, Hugo; Van Paegem, Wim

    2014-01-01

    A method is introduced for the measurements of strain exploiting the interaction between ultrasound waves and characteristics of the insonified specimen. First, the response of obliquely incident harmonic waves to a deterministic surface roughness is utilized. Analysis of backscattered amplitudes in Bragg diffraction geometry then yields a measure for the in-plane strain field by mapping any shift in angular dependency. Secondly, the analysis of the reflection characteristics of normal incide...

  10. Role of killer factors in the inhibitory activity of bio-control yeasts against Penicillium expansum and Aspergillus ochraceus

    OpenAIRE

    Ciro da Silva Portes; Andriélen Virke de Oliveira; Patrícia Simer; Alessandra Machado Lunkes; Alexandre Rodrigo Coelho

    2013-01-01

    This work evaluated the antagonism of killer positive yeast strains (isolated from 11 samples of different frozen fruit pulps) against the strains of Penicillium expansum and Aspergillus ochraceus. Of the total 41 killer yeasts tested in YM agar, 19 showed antibiosis against P. expansum and A. ochraceus, with inhibition zone ranging from 10 to 18 mm and 10 to 19 mm, respectively. In the following step, the extracellular activity of Kluyveromyces sp. FP4(13) was tested performing the assay in ...

  11. 生物防治白僵菌与家蚕病原白僵菌的生物学特性初步比较%A Preliminary Comparison on Biological Characteristics of Biocontrol Agent Beauveria bassiana and Silkworm Pathogen Beauveria bassiana

    Institute of Scientific and Technical Information of China (English)

    刘吉平; 吕思行; 米红霞

    2011-01-01

    In order to provide experimental data for the source-tracking and control of white muscardine disease and for safety assessment of applying Beauveria bassiana insecticide in silkworm rearing areas, PDA medium and scanning electron microscopy were used for isolation, identification and observation of 2 biocontrol agents, namely B. bassiana strain Bb2 used for forest protection in Guangzhou City and B. bassiana strain Bb7 preserved in the South China Agricultural University, and of 3 B. bassiana isolates collected in Guangdong provincial silkworm rearing areas at Huazhou Silkworm Eggs Production Farm, Daqiao Village of Huazhou City, and Xiao Village of Wengyuan County, respectively. A prelimina ry comparison was conducted on the biological characteristics between the above5 strains. The results showed that there existed differences on the colonial morphology, vegetative growth, spore quantity and other biological characteristics of the 5 strains. Among them, strain Bb2 grew the fastest, and strains Bb8 and Bb13 had the highest spore quantities. The conidiospores from muscardine pupae infected by strains Bb8 and Bb14 were spherical or near-spherical under scanning electron microscope. Both of them belonged to [ Beauveria bassiana (Bals.) Vuill. ] The results also showed that there existed certain similarities in biological characteristics between the pathogens of silkworm white muscardine from some silkworm rearing areas and from biocontrol agent B. bassiana. Meanwhile, some diversity also existed in B. bassiana strains from the same silkworm rearing area. Therefore, it is implied that the source of white muscardine is rather complex, and further investigation is thus needed to ascertain whether the two biocontrol agents Bb2 and Bb7 belong to B. bassiana.%对广州市森林保护用生防白僵菌Bb2、华南农业大学保存生防用白僵菌Bb7以及收集自广东省化州蚕种场、化州市笪桥村、翁源县硝村等蚕区的3株家蚕白僵菌分离株(Bb8

  12. Biocontrole da mancha-aquosa do melão pelo tratamento de sementes com bactérias epifíticas e endofíticas Biocontrol of bacterial blotch of melon by seed treatment with epiphytic and endophytic bacteria

    Directory of Open Access Journals (Sweden)

    Aldenir de Oliveira

    2006-09-01

    avenae subsp. citrulli or treating healthy seeds in order to protect the plant challenged later with the pathogen. Melon seeds were microbiolized by immersion in suspensions (A570= 0,7, sown and evaluated for incubation period (PI, incidence (INC, disease severity (SEV and disease severity reduction (RSD. Only microbiolization of artificially infected seed using the endophytic ENM5 (not identified, ENM9 (Bacillus cereus, ENM13 (Bacillus sp., ENM16 (Bacillus cereus, ENM32 (Bacillus subtilis and ENM43 (Bacillus sp., showed potencial for disease control. After in vitro compatibility tests, these strains were reevaluated separately and in pairs, in relation to PI, INC, SEV, RSD, disease index (IDO and area under disease progress curve (AACPD. All treatments differed significantly (P= 0.05 from the control reaching 93.6% of RSD, with emphasis for ENM13 and ENM9 with 7.5 and 7.25 days of PI, 0.22 and 0.22 of SEV, 2.59 and 2.59 of IDO, and 0.22 and 0.39 of AACPD, respectively. In vitro tests to determine the putative mechanisms of action involved in biocontrol were performed. ENM13 and ENM9 solubilized phosphate, ENM5 presented antibiose against A. avenae subsp. citrulli, and ENM43 produced HCN while ENM16 and ENM32 did not show any of the tested mechanisms.

  13. Biological Control of Bipolaris sorokiniana on Tall Fescue by Stenotrophomonas maltophilia Strain C3.

    Science.gov (United States)

    Zhang, Z; Yuen, G Y

    1999-09-01

    ABSTRACT Stenotrophomonas maltophilia strain C3 was evaluated for control of leaf spot on tall fescue (Festuca arundinacea) caused by Bipolaris sorokiniana. In growth chamber experiments, C3 inhibited conidial germination on leaf surfaces and reduced lesion frequency and percent diseased leaf area compared with nontreated controls. The amount of leaf spot suppression was related to the C3 dose applied. The highest dose tested, 10(9) CFU/ml, prevented nearly all B. sorokiniana conidia from germinating on treated leaf surfaces and provided nearly complete suppression of lesion development. When colloidal chitin was added to C3 cell suspensions of 10(7) or 10(8) CFU/ml, biocontrol efficacy was significantly increased over C3 applied alone, whereas addition of chitin to a C3 cell suspension of 10(9) CFU/ml had no effect. In field experiments, application of C3 to tall fescue turf resulted in significant reductions in infection frequency and disease severity compared with nontreated controls. Strain C3 applied at 10(9) CFU/ml was more effective than C3 applied at 10(7) CFU/ml, and amendment of the lower dose with colloidal chitin enhanced its efficacy. Populations sizes of C3 established on foliage in a growth chamber and in the field were directly related to dose applied. Chitin amendments did not affect C3 population size.

  14. Screening,Identification and Biocontrol Potential of Antagonistic Fungi Against Strawberry Root Rot and Plant Growth Promotion%草莓根腐病拮抗真菌筛选鉴定及其防病促生作用

    Institute of Scientific and Technical Information of China (English)

    申光辉; 薛泉宏; 张晶; 段佳丽; 王东胜; 杨兴华

    2012-01-01

    [目的]探索利用植物病、健株根域土壤微生物区系比较法快速筛选高效、广谱且有实际防病促生作用生防真菌的可行性.[方法]根据菌落培养与扫描电镜形态特征及rDNA-ITS序列对拮抗真菌进行鉴定;生长速率法测定菌株无菌发酵滤液的拮抗活性,盖玻片对峙培养法观察拮抗真菌对草莓根腐病原菌菌丝形态的影响;室内发芽试验法测定真菌无菌发酵滤液促种子萌发活性;采用盆栽试验研究拮抗真菌对草莓根腐病的防病促生作用;土壤稀释平皿分离法测定拮抗真菌的定殖能力及对草莓根域微生物区系的影响.[结果]经鉴定,2株拮抗真菌HF3和HF7分别为灰黄青霉(Penicillium griseofulvum)和土曲霉(Aspergillus terreus),其无菌发酵滤液对7种供试病原真菌有较强的拮抗作用,培养96 h时,HF3对立枯丝核菌(Rhizoctonia solani)和大丽轮枝菌(Verticillium dahliae)菌丝生长抑菌率均为100.0%.HF3菌丝对大双孢柱孢(Cylindrocarpon macrodidyma CF9)菌丝具有溶解作用,使病菌菌丝出现扭曲、变细、断裂及原生质浓缩等畸变现象;HF3和HF7 103和104倍稀释无菌发酵滤液可促进甜瓜种子萌发及胚根、胚芽生长.HF7 104倍稀释液对西瓜种子萌发及胚芽、胚根生长具有促进作用;HF3和HF7对盆栽草莓根腐病的防效分别达53.0%和46.9%,同时可促进盆栽草莓植株生长,提高果实产量;菌株HF3和HF7在草莓根表定殖数量分别为2.23×105、1.02×105 CFU·g-1干土,同时减少了大双孢柱孢数量;接种HF3和HF7可有效调整草莓根域微生物区系,促进土壤微生态恢复平衡.[结论]通过植物病、健株根区土壤微生物区系比较可以快速筛选到高效广谱且有实际防病促生作用的生防真菌;HF3和HF7对草莓根腐病防治具有较高的潜在应用价值.%[Objective] The objective of this study is to verify the suitability of a rapid screening program for biocontrol

  15. Typhlodromus pyri and Euseius finlandicus (Acari: Phytoseiidae) as potential biocontrol agents against spider mites (Acari: Tetranychidae) inhabiting willows: laboratory studies on predator development and reproduction on four diets.

    Science.gov (United States)

    Puchalska, Ewa K; Kozak, Marcin

    2016-01-01

    Typhlodromus pyri Scheuten and Euseius finlandicus (Oudemans) are important predators of phytophagous mites. The present laboratory study aimed to determine whether both species can develop and reach maturity feeding on spider mites occurring on willows, i.e., Schizotetranychus schizopus (Zacher), Schizotetranychus garmani Pritchard & Baker, and Tetranychus urticae Koch, and on Brassica napus L. pollen. The predators' development, reproduction and demographic parameters were significantly affected by diet. The data suggest that rape pollen can be useful in mass rearing of E. finlandicus but is completely unsuitable as alternative food for T. pyri. Short development time and high values of population parameters achieved by T. pyri feeding on larvae and protonymphs of S. schizopus and by E. finlandicus feeding on juvenile stages of S. garmani indicate great suitability of these preys as food for the phytoseiids, and make both predatory species promising biocontrol agents in spider mite control on willows. PMID:26530991

  16. Typhlodromus pyri and Euseius finlandicus (Acari: Phytoseiidae) as potential biocontrol agents against spider mites (Acari: Tetranychidae) inhabiting willows: laboratory studies on predator development and reproduction on four diets.

    Science.gov (United States)

    Puchalska, Ewa K; Kozak, Marcin

    2016-01-01

    Typhlodromus pyri Scheuten and Euseius finlandicus (Oudemans) are important predators of phytophagous mites. The present laboratory study aimed to determine whether both species can develop and reach maturity feeding on spider mites occurring on willows, i.e., Schizotetranychus schizopus (Zacher), Schizotetranychus garmani Pritchard & Baker, and Tetranychus urticae Koch, and on Brassica napus L. pollen. The predators' development, reproduction and demographic parameters were significantly affected by diet. The data suggest that rape pollen can be useful in mass rearing of E. finlandicus but is completely unsuitable as alternative food for T. pyri. Short development time and high values of population parameters achieved by T. pyri feeding on larvae and protonymphs of S. schizopus and by E. finlandicus feeding on juvenile stages of S. garmani indicate great suitability of these preys as food for the phytoseiids, and make both predatory species promising biocontrol agents in spider mite control on willows.

  17. Muscle strain injuries.

    Science.gov (United States)

    Garrett, W E

    1996-01-01

    One of the most common injuries seen in the office of the practicing physician is the muscle strain. Until recently, little data were available on the basic science and clinical application of this basic science for the treatment and prevention of muscle strains. Studies in the last 10 years represent action taken on the direction of investigation into muscle strain injuries from the laboratory and clinical fronts. Findings from the laboratory indicate that certain muscles are susceptible to strain injury (muscles that cross multiple joints or have complex architecture). These muscles have a strain threshold for both passive and active injury. Strain injury is not the result of muscle contraction alone, rather, strains are the result of excessive stretch or stretch while the muscle is being activated. When the muscle tears, the damage is localized very near the muscle-tendon junction. After injury, the muscle is weaker and at risk for further injury. The force output of the muscle returns over the following days as the muscle undertakes a predictable progression toward tissue healing. Current imaging studies have been used clinically to document the site of injury to the muscle-tendon junction. The commonly injured muscles have been described and include the hamstring, the rectus femoris, gastrocnemius, and adductor longus muscles. Injuries inconsistent with involvement of a single muscle-tendon junction proved to be at tendinous origins rather than within the muscle belly. Important information has also been provided regarding injuries with poor prognosis, which are potentially repairable surgically, including injuries to the rectus femoris muscle, the hamstring origin, and the abdominal wall. Data important to the management of common muscle injuries have been published. The risks of reinjury have been documented. The early efficacy and potential for long-term risks of nonsteroidal antiinflammatory agents have been shown. New data can also be applied to the field

  18. Genetic diversity of Trichoderma atroviride strains collected in Poland and identification of loci useful in detection of within-species diversity.

    Science.gov (United States)

    Skoneczny, Dominik; Oskiera, Michał; Szczech, Magdalena; Bartoszewski, Grzegorz

    2015-07-01

    Molecular markers that enable monitoring of fungi in their natural environment or assist in the identification of specific strains would facilitate Trichoderma utilization, particularly as an agricultural biocontrol agent (BCA). In this study, sequence analysis of internal transcribed spacer regions 1 and 2 (ITS1 and ITS2) of the ribosomal RNA (rRNA) gene cluster, a fragment of the translation elongation factor 1-alpha (tef1) gene, and random amplified polymorphic DNA (RAPD) markers were applied to determine the genetic diversity of Trichoderma atroviride strains collected in Poland, and also in order to identify loci and PCR-based molecular markers useful in genetic variation assessment of that fungus. Although tef1 and RAPD analysis showed limited genetic diversity among T. atroviride strains collected in Poland, it was possible to distinguish major groups that clustered most of the analyzed strains. Polymorphic RAPD amplicons were cloned and sequenced, yielding sequences representing 13 T. atroviride loci. Based on these sequences, a set of PCR-based markers specific to T. atroviride was developed and examined. Three cleaved amplified polymorphic sequence (CAPS) markers could assist in distinguishing T. atroviride strains. The genomic regions identified may be useful for further exploration and development of more precise markers suitable for T. atroviride identification and monitoring, especially in environmental samples.

  19. Colonization and degradation of senescent flowers of zucchini squash by Trichoderma harzianum YC459, a biocontrol agent of gray mold, Botrytis cinerea

    Institute of Scientific and Technical Information of China (English)

    Geun Gon Kim; Young Ryun Chung

    2004-01-01

    @@ In commercial greenhouses, senescent flower petals or flowers of vegetables such as tomato,strawberry, hot pepper and zucchini squash were blighted to be removed from fruits within five days after spraying of Trichoderna harzianun YC459 (TORY() , JGreen Inc.), a biocontrol agent with good and consistent efficacy as chemical fungicides for the control of gray mold rot caused by B.cinerea. The mechanism for selective colonization of senescent floral tissues by T. harzianum YCA59was elucidated using fresh and senescent (4 days and 14 days after pollination, respectively) floral tissues of zucchini squash (Cucurbita moschata Duchesne) . The spores of T. harzianum YCA59were produced much more on water agar and liquid culture media supplemented with 5% dry powder of senescent floral tissues than with fresh tissues during 15 days incubation. Mycelial growth was also much better in the media with senescent tissues than with fresh tissues. Enzyme activities of carboxymethyl cellulase, amylase and polygalacturonase in the liquid media, which might be involved in the colonization and degradation of tissues by T. harzianum YCA59 were compared. The activities of three enzymes were significantly higher in the media with senescent floral tissues than with fresh floral tissues reaching to the maximum during 9 to 12 days of incubation. Especially, the activities of carboxymethyl cellulase and polygalacturonase of T. harzianum YC459 were much higher than those of other Trichoderma species, T. asperellum, T. viride and T. koningii in the liquid media with senescent floral tissues. Based on the results, the selective colonization and degradation of senescent floral tissues, an important habitat for B. cinerea, may be another mechanism for the biocontrol of gray mold rot of vegetables by T. harzianurm YC459.

  20. In Vitro Evaluation Of Selected Plant Extracts As Biocontrol Agents Against Black Mold Aspergillus Niger Van Tieghem Of Onion Bulbs Allium Cepa L.

    Directory of Open Access Journals (Sweden)

    Saifeldin A. F. El-Nagerabi

    2015-08-01

    Full Text Available Black mold disease caused by Aspergillus niger V. Tiegh. is the most devastating infection occurs in onions Allium cepa L. under field and store conditions. The use of biocontrol agents is ecofriendly approach for controlling seedborne and soilborne diseases compared to the use of toxic synthetic fungicides. This study has been designed to assess the contamination levels of onion seeds with A. niger and its effect on seed germination and to evaluate the in vitro antifungal activity of Prunus mahaleb seeds Commiphora myrrha resin 0.5 1.0 1.5 2.0 g100 ml Syzygium aromaticum dry buds clove and Panax ginseng roots extracts 0.5 1.0 2.0 2.5 g100 ml against black mold of onion bulbs. The fungus is seedborne pathogen which significantly contaminated onion seeds 89-100 and reduced seed germination 39-83. The extracts of clove caused 43-96 inhibition in spore germination followed by mahaleb 37-96 myrrha 33-88 and ginseng 34-87. The highest concentration of these extracts 3.0 did not affect seed germination but significantly reduced seed contamination by A. niger up to 84 80 71 and 65 for Syzygium aromaticum Syzygium aromaticum Panax ginseng and Prunus mahaleb respectively. The extracts apparently inhibited the fungal growth and mold development on stored onion bulbs which indicates the antifungal property of these extracts against A. niger. Therefore they can be recommended as effective biocontrol agents to reduce seed contamination and enhance the storability of onion bulbs. Thus the use of healthy and certified seeds for onion production is a priority.

  1. The Pochonia chlamydosporia Serine Protease Gene vcp1 Is Subject to Regulation by Carbon, Nitrogen and pH: Implications for Nematode Biocontrol

    Science.gov (United States)

    Ward, Elaine; Kerry, Brian R.; Manzanilla-López, Rosa H.; Mutua, Gerald; Devonshire, Jean; Kimenju, John; Hirsch, Penny R.

    2012-01-01

    The alkaline serine protease VCP1 of the fungus Pochonia chlamydosporia belongs to a family of subtilisin-like enzymes that are involved in infection of nematode and insect hosts. It is involved early in the infection process, removing the outer proteinaceous vitelline membrane of nematode eggs. Little is known about the regulation of this gene, even though an understanding of how nutrients and other factors affect its expression is critical for ensuring its efficacy as a biocontrol agent. This paper provides new information on the regulation of vcp1 expression. Sequence analysis of the upstream regulatory region of this gene in 30 isolates revealed that it was highly conserved and contained sequence motifs characteristic of genes that are subject to carbon, nitrogen and pH-regulation. Expression studies, monitoring enzyme activity and mRNA, confirmed that these factors affect VCP1 production. As expected, glucose reduced VCP1 expression and for a few hours so did ammonium chloride. Surprisingly, however, by 24 h VCP1 levels were increased in the presence of ammonium chloride for most isolates. Ambient pH also regulated VCP1 expression, with most isolates producing more VCP1 under alkaline conditions. There were some differences in the response of one isolate with a distinctive upstream sequence including a variant regulatory-motif profile. Cryo-scanning electron microscopy studies indicated that the presence of nematode eggs stimulates VCP1 production by P. chlamydosporia, but only where the two are in close contact. Overall, the results indicate that readily-metabolisable carbon sources and unfavourable pH in the rhizosphere/egg-mass environment may compromise nematode parasitism by P. chlamydosporia. However, contrary to previous indications using other nematophagous and entomopathogenic fungi, ammonium nitrate (e.g. from fertilizers) may enhance biocontrol potential in some circumstances. PMID:22558192

  2. The Pochonia chlamydosporia serine protease gene vcp1 is subject to regulation by carbon, nitrogen and pH: implications for nematode biocontrol.

    Directory of Open Access Journals (Sweden)

    Elaine Ward

    Full Text Available The alkaline serine protease VCP1 of the fungus Pochonia chlamydosporia belongs to a family of subtilisin-like enzymes that are involved in infection of nematode and insect hosts. It is involved early in the infection process, removing the outer proteinaceous vitelline membrane of nematode eggs. Little is known about the regulation of this gene, even though an understanding of how nutrients and other factors affect its expression is critical for ensuring its efficacy as a biocontrol agent. This paper provides new information on the regulation of vcp1 expression. Sequence analysis of the upstream regulatory region of this gene in 30 isolates revealed that it was highly conserved and contained sequence motifs characteristic of genes that are subject to carbon, nitrogen and pH-regulation. Expression studies, monitoring enzyme activity and mRNA, confirmed that these factors affect VCP1 production. As expected, glucose reduced VCP1 expression and for a few hours so did ammonium chloride. Surprisingly, however, by 24 h VCP1 levels were increased in the presence of ammonium chloride for most isolates. Ambient pH also regulated VCP1 expression, with most isolates producing more VCP1 under alkaline conditions. There were some differences in the response of one isolate with a distinctive upstream sequence including a variant regulatory-motif profile. Cryo-scanning electron microscopy studies indicated that the presence of nematode eggs stimulates VCP1 production by P. chlamydosporia, but only where the two are in close contact. Overall, the results indicate that readily-metabolisable carbon sources and unfavourable pH in the rhizosphere/egg-mass environment may compromise nematode parasitism by P. chlamydosporia. However, contrary to previous indications using other nematophagous and entomopathogenic fungi, ammonium nitrate (e.g. from fertilizers may enhance biocontrol potential in some circumstances.

  3. Biocontrol of Botrytis allii Munn the Causal Agent of Neck Rot, the Post Harvest Disease in Onion, by use of a New Iranian Isolate of Streptomyces

    Directory of Open Access Journals (Sweden)

    M. Jorjandi

    2009-01-01

    Full Text Available Problem statement: Soil actinomycetes particularly Streptomyces spp. showed antagonistic activity against wide range of plant pathogens. In the recent decades they have attracted high interests as biocontrol agents. Onion neck rot or gray mold caused by Botrytis allii have imposed economic post harvest damages to onion bulbs and decreased its storage durability and market value. Approach: To investigate for biocontrol means against the pathogen, antagonistic activity of 50 isolates of soil Actinomycetes were assayed through agar disk method and dual culture bioassays. Active isolates were exposed to chloroform for detection of antibiotic. Minimum Inhibitory Concentration (MIC value and solubility of active crude extract in organic solvents were determined for Streptomyces isolate No. 347 which showed a unique and stable property of inhibiting Botrytis allii. To investigate the antagonistic effect of Streptomyces isolate No. 347 on control of onion gray mold, 4 different treatments were tested by means of Tukey HSD test. Results: From the tested isolates, 13 showed anti gray mold activities. Exposure of active isolates to chloroform revealed that Streptomyces isolates No. 347, 263 and 350 retained their antifungal activities. The active metabolite(s of Streptomyces isolate No. 347 was polar, soluble in H2O but insoluble in chloroform and methanol. MIC of the crude was determined as 0.05 mg mL-1 against B. allii. Stability of the active crude in distilled water at room temperature (12-30°C was about 6 months. Statistical studies indicated that Streptomyces isolates No. 347 can decrease losses of neck rot with significant level (pConclusion: The future goals include investigation of the antifungal genes in active isolates as candidates for genetic engineering of onion for increased tolerance against B. allii.

  4. Longitudinal misalignment based strain sensor

    OpenAIRE

    Andrews, Jeffrey Pratt

    1989-01-01

    A practical fiber optic strain sensor has been developed to measure strains in the range of 0.0 to 2.0 percent strain with a resolution ranging between 10 and 100 microstrain depending on sensor design choices. This intensity based sensor measures strain by monitoring strain induced longitudinal misalignment in a novel fiber interconnection. This interconnection is created by aligning fibers within a segment of hollow core fiber. Related splice loss mechanisms are investigated ...

  5. Expression of cry1Ab gene from a novel Bacillus thuringiensis strain SY49-1 active on pest insects.

    Science.gov (United States)

    Azizoglu, Ugur; Ayvaz, Abdurrahman; Yılmaz, Semih; Karabörklü, Salih; Temizgul, Rıdvan

    2016-01-01

    In this study, the cry1Ab gene of previously characterized and Lepidoptera-, Diptera-, and Coleoptera-active Bacillus thuringiensis SY49-1 strain was cloned, expressed and individually tested on Ephestia kuehniella (Lepidoptera: Pyralidae) and Plodia interpunctella (Lepidoptera: Pyralidae) larvae. pET-cry1Ab plasmids were constructed by ligating the cry1Ab into pET28a (+) expression vector. Constructed plasmids were transferred to an Escherichia coli BL21 (DE3) strain rendered competent with CaCl2. Isopropyl β-d-1-thiogalactopyranoside was used to induce the expression of cry1Ab in E. coli BL21(DE3), and consequently, ∼130kDa of Cry1Ab was obtained. Bioassay results indicated that recombinant Cry1Ab at a dose of 1000μgg(-1) caused 40% and 64% mortality on P. interpunctella and E. kuehniella larvae, respectively. However, the mortality rates of Bt SY49-1 strains' spore-crystal mixture at the same dose were observed to be 70% on P. interpunctella and 90% on E. kuehniella larvae. The results indicated that cry1Ab may be considered as a good candidate in transgenic crop production and as an alternative biocontrol agent in controlling stored product moths.

  6. A Saccharomyces cerevisiae Wine Strain Inhibits Growth and Decreases Ochratoxin A Biosynthesis by Aspergillus carbonarius and Aspergillus ochraceus

    Directory of Open Access Journals (Sweden)

    Marilena Budroni

    2012-12-01

    Full Text Available The aim of this study was to select wine yeast strains as biocontrol agents against fungal contaminants responsible for the accumulation of ochratoxin A (OTA in grape and wine and to dissect the mechanism of OTA detoxification by a Saccharomyces cerevisiae strain (DISAABA1182, which had previously been reported to reduce OTA in a synthetic must. All of the yeast strains tested displayed an ability to inhibit the growth of Aspergillus carbonarius both in vivo and in vitro and addition of culture filtrates from the tested isolates led to complete inhibition of OTA production. S. cerevisiae DISAABA1182 was selected and further tested for its capacity to inhibit OTA production and pks (polyketide synthase transcription in A. carbonarius and Aspergillus ochraceus in vitro. In order to dissect the mechanism of OTA detoxification, each of these two fungi was co-cultured with living yeast cells exposed to yeast crude or to autoclaved supernatant: S. cerevisiae DISAABA1182 was found to inhibit mycelial growth and OTA production in both Aspergilli when co-cultured in the OTA-inducing YES medium. Moreover, a decrease in pks transcription was observed in the presence of living cells of S. cerevisiae DISAABA1182 or its supernatant, while no effects were observed on transcription of either of the constitutively expressed calmodulin and β-tubulin genes. This suggests that transcriptional regulation of OTA biosynthetic genes takes place during the interaction between DISAABA1182 and OTA-producing Aspergilli.

  7. Expression of cry1Ab gene from a novel Bacillus thuringiensis strain SY49-1 active on pest insects.

    Science.gov (United States)

    Azizoglu, Ugur; Ayvaz, Abdurrahman; Yılmaz, Semih; Karabörklü, Salih; Temizgul, Rıdvan

    2016-01-01

    In this study, the cry1Ab gene of previously characterized and Lepidoptera-, Diptera-, and Coleoptera-active Bacillus thuringiensis SY49-1 strain was cloned, expressed and individually tested on Ephestia kuehniella (Lepidoptera: Pyralidae) and Plodia interpunctella (Lepidoptera: Pyralidae) larvae. pET-cry1Ab plasmids were constructed by ligating the cry1Ab into pET28a (+) expression vector. Constructed plasmids were transferred to an Escherichia coli BL21 (DE3) strain rendered competent with CaCl2. Isopropyl β-d-1-thiogalactopyranoside was used to induce the expression of cry1Ab in E. coli BL21(DE3), and consequently, ∼130kDa of Cry1Ab was obtained. Bioassay results indicated that recombinant Cry1Ab at a dose of 1000μgg(-1) caused 40% and 64% mortality on P. interpunctella and E. kuehniella larvae, respectively. However, the mortality rates of Bt SY49-1 strains' spore-crystal mixture at the same dose were observed to be 70% on P. interpunctella and 90% on E. kuehniella larvae. The results indicated that cry1Ab may be considered as a good candidate in transgenic crop production and as an alternative biocontrol agent in controlling stored product moths. PMID:27143037

  8. Correlation between virulence and genetic structure of Escovopsis strains from leaf-cutting ant colonies in Costa Rica.

    Science.gov (United States)

    Wallace, Diego E Elizondo; Asensio, Juan G Vargas; Tomás, Adrián A Pinto

    2014-08-01

    Leaf-cutting ants (genera Atta and Acromyrmex) cultivate a specialized fungus for food in underground chambers employing cut plant material as substrate. Parasitism occurs in this agricultural system and plays an important role in colony fitness. The microfungi Escovopsis, a specialized mycoparasite of the fungal cultivar, is highly prevalent among colonies. In this study, we tested the antagonistic activity of several Escovopsis strains from different geographical areas in Costa Rica. We employed a combination of laboratory tests to evaluate virulence, including pure culture challenges, toxicity to fungus garden pieces and subcolony bioassays. We also performed a phylogenetic analysis of these strains in order to correlate their virulence with the genetic structure of this population. The bioassays yielded results consistent between each other and showed significant differences in antagonistic activity among the parasites evaluated. However, no significant differences were found when comparing the results of the bioassays according to the source of the ants' fungal cultivar. The phylogenetic analyses were consistent with these results: whilst the fungal cultivar phylogeny showed a single clade with limited molecular variation, the Escovopsis phylogeny yielded several clades with the most virulent isolates grouping in the same well-supported clade. These results indicate that there are Escovopsis strains better suited to establish their antagonistic effect, whilst the genetic homogeneity of the fungal cultivars limits their ability to modulate Escovopsis antagonism. These findings should be taken into consideration when evaluating the potential of Escovopsis isolates as biocontrol agents for this important agricultural pest in the Neotropics. PMID:24836623

  9. Asymptomatic bacteriuria Escherichia coli strains

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Nielsen, E.M.; Klemm, Per

    2006-01-01

    to uropathogenic E. coli (UPEC) that cause symptomatic UTI, very little is known about the mechanisms by which these strains colonize the urinary tract. Here, we have investigated the growth characteristics in human urine as well as adhesin repertoire of nine ABU strains; the ability of ABU strains to compete...... against the UPEC strain CFT073 was also studied. The different ABU strains displayed a wide variety of the measured characteristics. Half of the ABU strains displayed functional type 1 fimbriae while only one expressed functional P fimbriae. A good correlation between the growth rate of a particular...

  10. 新疆骆驼刺拮抗内生细菌XJAS-AB-13的分选·鉴定及其对玉米斑点病菌的生物防治潜能研究%Screening and Identification of An Antagonistic Endobacterium ( XJAS-AB-13 ) from Xinjiang Alhagi pseudalhagi Desv and Studies on Its Biocontrol Potentials to Maize Spot Pathogens

    Institute of Scientific and Technical Information of China (English)

    阿布都卡地尔·阿布力孜; 吾甫尔·米吉提; 阿依乃再·孜亚克; 阿力木江·阿合幼力

    2011-01-01

    [目的] 从骆驼刺(Alhagi pseudalhagi Desv)内生菌中筛选出对农作物致病菌具有较强生物活性的拮抗菌株,活体验证拮抗菌株的生物防治能力.[方法] 采用琼脂扩散法对常见的农作物致病菌做拮抗试验,结合传统病情统计和不同处理组玉米叶片中总蛋白含量以及防御酶活性测定活体验证拮抗菌株的生防能力.[结果] 筛选出对玉米大、小斑点病菌具有较强活性拮抗菌株XJAS-AB-13,根据拮抗内生细菌XJAS-AB-13的生理生化特征以及16S rDNA序列分析将其最终鉴定为枯草芽孢杆菌.该菌株发酵液对玉米大斑病菌和玉米小班病菌的抑菌圈直径分别达32.8和24.6 mm.活体试验结果显示,发酵液对玉米小斑病菌和玉米大斑病菌的防效率分别达到45.00%和63.33%.[结论] 拮抗菌株XJAS-AB-13在玉米大、小斑点病菌的生物防治中具有较好的应用潜力.%[ Objective ] The aim was to screen antagonistic endophytes from Alhagi pseudalhagi Desv to common crop pathogens, and verify biocontrol capacity of the active strains with in vivo experiment. [ Method] Agar diffusion method was used in screening the antagonistic endophytes. Traditional disease severity investigating method, measuring the total protein contents and defense enzymes activity of the maize leaf samples under the different treatment were used in testing the in vivo activity of the antagonistic strain. [Result] An antagonistic endobacterium (XJAS-AB-13) with strong antifungal activity to Exserohilum turcicum and Bipolaris maydis, was screened from Alhagi pseudalhagi Desv XJAS-AB-13 was finally identified as Bacillus subtilis according to physiological and biochemical characteristics and 16S rDNA sequence features. Inhibition zone diameter of fermentation broth of XJAS-AB-13 to Exserohilum turcicum and Bipolaris maydis reached 32. 8 and 24. 6 ram, respectively, in vivo experiment results showed that the biocontrol rate of XJAS-AB-13 to

  11. Strains in General Relativity

    CERN Document Server

    Bini, Donato; Geralico, Andrea

    2014-01-01

    The definition of relative accelerations and strains among a set of comoving particles is studied in connection with the geometric properties of the frame adapted to a "fiducial observer." We find that a relativistically complete and correct definition of strains must take into account the transport law of the chosen spatial triad along the observer's congruence. We use special congruences of (accelerated) test particles in some familiar spacetimes to elucidate such a point. The celebrated idea of Szekeres' compass of inertia, arising when studying geodesic deviation among a set of free-falling particles, is here generalized to the case of accelerated particles. In doing so we have naturally contributed to the theory of relativistic gravity gradiometer. Moreover, our analysis was made in an observer-dependent form, a fact that would be very useful when thinking about general relativistic tests on space stations orbiting compact objects like black holes and also in other interesting gravitational situations.

  12. Strains in general relativity

    International Nuclear Information System (INIS)

    The definition of relative accelerations and strains among a set of comoving particles is studied in connection with the geometric properties of the frame adapted to a 'fiducial observer'. We find that a relativistically complete and correct definition of strains must take into account the transport law of the chosen spatial triad along the observer's congruence. We use special congruences of (accelerated) test particles in some familiar spacetimes to elucidate such a point. The celebrated idea of Szekeres' compass of inertia, arising when studying geodesic deviation among a set of free-falling particles, is here generalized to the case of accelerated particles. In doing so we have naturally contributed to the theory of relativistic gravity gradiometer. Moreover, our analysis was made in an observer-dependent form, a fact that would be very useful when thinking about general relativistic tests on space stations orbiting compact objects like black holes and also in other interesting gravitational situations

  13. 生防菌解淀粉芽孢杆菌抗菌蛋白的研究进展%Research Advances of Antifungal Proteins Produced by Biocontrol Bacterium,Bacillus amyloliquefaciens

    Institute of Scientific and Technical Information of China (English)

    出晓铭; 林毅雄; 张珅; 严芬; 林河通

    2014-01-01

    Bacillus amyloliquefaciens is a kind of biocontrol bacterium which has great ability against fungi and bacteria,thereby it has high research values and potential for development in biological control of posthar-vest diseases of fruits and vegetables.The separation and purification of antifungal proteins produced by Bacil-lus amyloliquefaciens,the disease resistant mechanisms and the application of biocontrol of antifungal proteins were summarized.The future potential application in biocontrol of postharvest diseases of fruits and vegetables was also discussed.%生防解淀粉芽孢杆菌具有强烈抑制真菌和细菌的能力,在果蔬采后病害防治方面具有巨大的应用潜力。该文对解淀粉芽孢杆菌产生的抗菌蛋白、蛋白的分离纯化、抑菌机理和生防应用等方面进行了综述,并对其在果蔬采后病害防治应用前景进行了展望。

  14. MEMS Resonant Strain Sensor Integration

    OpenAIRE

    Myers, David Richard

    2010-01-01

    Despite commercial availability since the 1950's, silicon strain sensors have not experienced the same success as other microdevices, such as accelerometers, pressure sensors, and inkjet heads. Strain sensors measure mechanical deformation and could be used in many structural components, improving safety, controls, and manufacturing tolerances. This thesis examines major strain sensing techniques and highlights both advantages and disadvantages of each. MEMS resonant strain gauges are iden...

  15. Mutagenesis of beta-1,3-Glucanase Genes in Lysobacter enzymogenes Strain C3 Results in Reduced Biological Control Activity Toward Bipolaris Leaf Spot of Tall Fescue and Pythium Damping-Off of Sugar Beet.

    Science.gov (United States)

    Palumbo, Jeffrey D; Yuen, Gary Y; Jochum, C Christine; Tatum, Kristin; Kobayashi, Donald Y

    2005-06-01

    ABSTRACT Lysobacter enzymogenes produces extracellular lytic enzymes capable of degrading the cell walls of fungi and oomycetes. Many of these enzymes, including beta-1,3-glucanases, are thought to contribute to the biological control activity expressed by several strains of the species. L. enzymogenes strain C3 produces multiple extracellular beta-1,3-glucanases encoded by the gluA, gluB, and gluC genes. Analysis of the genes indicates they are homologous to previously characterized genes in the related strain N4-7, each sharing >95% amino acid sequence identity to their respective counterparts. The gluA and gluC gene products encode enzymes belonging to family 16 glycosyl hydrolases, whereas gluB encodes an enzyme belonging to family 64. Mutational analysis indicated that the three genes accounted for the total beta-1,3-glucanase activity detected in culture. Strain G123, mutated in all three glucanase genes, was reduced in its ability to grow in a minimal medium containing laminarin as a sole carbon source. Although strain G123 was not affected in antimicrobial activity toward Bipolaris sorokiniana or Pythium ultimum var. ultimum using in vitro assays, it was significantly reduced in biological control activity against Bipolaris leaf spot of tall fescue and Pythium damping-off of sugar beet. These results provide direct supportive evidence for the role of beta-1,3-glucanases in biocontrol activity of L. enzymogenes strain C3.

  16. Development of reversible strain gage

    International Nuclear Information System (INIS)

    A high-temperature strain gage which can be peeled after taking required apparent strain measurements in a furnace and can be attached reverse-side-up at the point of interest on a test structure was developed. Using the ''reversible'' strain gage with selected room-temperature curing type polyester adhesive, one can expect to measure thermal strain accurately, especially for on large structures, at the first test in temperature up to 250 deg C. The repeatability of apparent strains for about 100 reversible gages was within 50 microstrains of difference at 250 deg C (within 30 microstrains of difference for 80 % of the test gages). (author)

  17. [Echinococcus and strain concepts].

    Science.gov (United States)

    Utük, Armağan Erdem; Simsek, Sami

    2008-01-01

    Hydatid disease (echinococcosis) is one of the most important parasitic zoonoses and remains a public health and economic problem all over the world. Echinococcus granulosus includes a number of genetic variants and, up to date, analyses of mitochondrial DNA sequences have identified ten distinct genetic types (genotypes G1-10). This categorization follows closely the pattern of strain variation emerging based on biological characteristics. The extensive variation in E. granulosus may influence life-cycle patterns, host specificity, development rate, antigenicity, transmission dynamics, sensitivity to chemotherapeutic agents, and pathology. In this review, the recent genetic characterizations of Echinococcus genus have been summarized. PMID:18351549

  18. IN VITRO MANAGEMENT OF CURVULARIA LEAF SPOT OF MAIZE USING BOTANICALS, ESSENTIAL OILS AND BIO-CONTROL AGENTS

    OpenAIRE

    JYOTIKA PUROHIT; SRINIVASANRAGHVAN, A.; PRADEEP KUMAR; SUNAINA BISHT

    2013-01-01

    Different plant extracts, essential oils and different strains of Trichoderma harzianum were evaluated in vitroagainst Curvularia lunata. Amongst the plant extracts, Lantana was highly effective @ 15 per cent (86.76 inhibition%) and 20 per cent (89.49 inhibition %) followed by Morphankhi @ 5 per cent (83.53 %) and 10 per cent (85.88%) respectively. Among the essential oils, complete inhibition was recorded in Citronella oil at all 3 concentrations(2µL, 4µL and 8) and Peppermint oil at 4µL and...

  19. Reconstruction of axisymmetric strain distributions via neutron strain tomography

    International Nuclear Information System (INIS)

    Predicting the behaviour of structural components under a particular set of loading conditions requires knowledge of the residual elastic strain distribution throughout the bulk of these components. Characterising the 3D strain state at any particular point involves the measurement of six independent components which make up the second order strain tensor. Mapping the complete strain distribution throughout large volumes thus presents significant practical challenges. One possible solution to this problem is to reconstruct the 3D variation of strain components using tomographic techniques. The basic principle underpinning this idea is that the multi-component strain tensor can be reconstructed from a redundant set of lower order projection data. Here we demonstrate this fundamental concept for two samples: a shrink fit ‘ring-and-plug’ sample, and a spray-quenched circular cylinder, both possessing axially symmetric internal strain distribution. We present and contrast different approaches to the strain tomography problem. The methods described here can also be readily applied to high-energy X-ray diffraction measurements and represent an important step toward developing the tomographic reconstruction framework for strain tensor distributions of arbitrary complexity. The major benefit of neutron strain tomography is that the incident beam flux is utilised more fully, greatly reducing the data collection times. Using micro-channel plate (MCP) neutron detectors, a spatial resolution of the order of 0.1 mm can be achieved .

  20. 贵州省黔东南州太子参立枯病的病害分析及生防初探%Study on Pathogen and Bio-control of Damping-off Disease in Pseudostellaria heterophylla in Qiandongnan Prefecture of Guizhou Province

    Institute of Scientific and Technical Information of China (English)

    张国辉; 王龙; 任永权; 贺定翔; 兰才武; 范成明; 陈宇红; 胡赞民

    2015-01-01

    Fungal diseases occur seriously in Pseudostel aria heterophyl a in Qian-dongnan Prefecture of Guizhou Province from March to May every year. By field in-vestigation, tissue isolation and microscopic identification, the main pathogen was iden-tified as Rhizoctonia solani, the pathogen of damping-off disease. At the same time, Alternaria alternate was also isolated from the sampled diseased soil, and it has pathogenicity to Pseudostel aria heterophyl a seedlings. The bio-control test showed that GH18 strain (Bacil us spp.) had significant inhibitory effects on Rhizoctonia solani and Alternaria alternate.%贵州省黔东南州太子参( Radix pseudostellariae)在每年3~5月份都会发生严重真菌病害,经实地调查、组织分离和显微鉴定,得到主要致病菌为立枯丝核菌(Rhizoctonia solani),其导致的病害为太子参立枯病。同时,还在病土中分离纯化得到对太子参种苗有致病能力的病原菌互隔文链孢霉(Alternariaalternate)。生防试验结果表明,芽孢杆菌(Bacillus spp.)GH18菌株对病原菌立枯丝核菌(Rhizoctoniasolani)和病原菌互隔文链孢霉(Alternaria alternate)的抑菌效果显著。